Identification of differentially expressed proteins involved in fetal scarless wound healing using a rat model of cleft lip
- Authors:
- Published online on: June 22, 2021 https://doi.org/10.3892/mmr.2021.12235
- Article Number: 596
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Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Abstract
Introduction
The cleft lip is a very common congenital oral and maxillofacial malformation, often accompanied by cleft palate and alveolar cleft. Although surgical repair techniques are continuously being improved, numerous patients still experience inevitable secondary scar formation after surgery. In recent years, with the development of prenatal diagnosis and treatment technology (1), intrauterine surgery has made it possible to correct developmental deformities, such as a cleft lip.
The concept of scarless healing was first proposed by Burrington (2) in 1971. It was later observed that fetal skin wounds that occur during early pregnancy can heal quickly and restore intact skin barrier functions. In contrast, fetal skin damage that occurs in the third trimester of pregnancy can result in the formation of scar tissue similar to that of an adult (3). Therefore, the different manifestations of scarless healing of mammalian fetal wounds are related to the gestational age of the fetus (4). Dang et al (5) and Longaker et al (6) demonstrated that this transition period from scarless healing to scar formation occurred between day 16.5 of gestational age (GA) and GA18.5 in rats and mice, which have a gestation period of ~21.5 days. Lorenz et al (7) and Cass et al (8) suggested that when 1–2 mm incisions are inflicted on fetal rats, the transition period of scarless healing to healing with scar formation was still between 16.5 (GA16.5) and 18.5 days (GA18.5).
This phenotypic difference in fetal wounds has inspired further examination of the specific underlying mechanisms. Initially, it was hypothesized that the reason for early scar repair was that the fetus developed in amniotic fluid, which is rich in growth factors and extracellular matrix (ECM) components (9,10). Previous studies typically utilized large animal models to study the presence of scars following repair (11,12). However, only a few studies have reported the use of a fetal rat cleft lip wound model to establish the effectiveness of surgical repair at different gestational ages. Moreover, due to the short gestation period of rats, the experimental cycle can be shortened, and the experiment can therefore be repeated.
Given the importance of this process, the present study, screened out several specific markers of early fetal scarless repair. The present study aimed to gain insight into the occurrence and mechanisms of scarless repair, and to identify new clinical targets for the prevention and treatment of scars.
Materials and methods
Animals
A total of 36 SPF-grade adult Sprague-Dawley (SD) rats (female; mean weight, 250 g; age, 12 weeks) were obtained from the Third Xiangya Hospital of Central South University Animal Experiment Center (Hunan, China) and divided into two groups that received surgery once their fetuses reached GA16.5 or GA18.5, respectively (n=18 each). The following housing conditions were implemented: A temperature between 25±2°C, relative humidity of 55±15%, ventilation rate of 10–20 times per hour, time-controlled artificial lighting (12-h day-night cycle) and ad libitum access to food and water. The experiments were supervised throughout and were performed in accordance with animal experimentation ethics.
Preliminary study on different repair modes applicable to fetal rats with artificial cleft lip wounds
Fetal rats located away from the uterine horn were selected to prevent subsequent abortion, as described previously (13). In the current study, rats were anesthetized with 30 mg/kg pentobarbital sodium intraperitoneally before surgery. A wedge-shaped cleft-like defect was created on the upper-left lip of the fetal rats. The upper-right lip did not receive any treatment and was used as a control condition. The fetal rats were then returned to the uterus. Fetal rats from the GA16.5 and GA18.5 groups were then removed three days post-surgery as previously described (4) (i.e., at GA19.5 or GA21.5, respectively). All fetuses and rats were euthanized using carbon dioxide (30% volume displaced/min). Death was confirmed using cervical dislocation. A total of three fetal rats were obtained from each pregnant rat, for a total of 54 fetal rats from both GA16.5 and GA18.5 groups, and the survival rate was calculated. Tissue samples from the surgical site on the upper-left lip and asymmetrical sections from the upper-right lip were collected from the fetal rats for histological examination, including hematoxylin and eosin (H&E) staining, Masson's Trichrome staining and type-I collagen immunohistochemical staining as previously described (14–16).
The upper-left lip tissue samples from the GA16.5 group were defined as group 1, whereas the upper-right lip tissue samples from the GA16.5 group were defined as group 2. In addition, the upper-left lip tissue samples from the GA18.5 group were defined as group 3, whereas the upper-right lip tissue samples from the GA18.5 group were defined as group 4. Each subgroup included 27 samples. Protein expression was compared between group 1 and 2, group 3 and 4, as well as group 3 and 1. Label-free quantification PRM was performed as previously described (17) and was used to detect the differentially expressed proteins among the different groups. MaxQuant 1.5.6 (https://www.maxquant.org) and Perseus 1.4 (https://www.maxquant.org/perseus/) were used to analyze the results of label-free quantification PRM: Volcano plots were generated for differentially expressed proteins: Y-axis, -log10(P-value); x-axis: log2(ratio). The points distributed outside the two vertical borders and above the horizontal border represented the proteins with significant differences; proteins with at least a 1.5-fold change in expression and P<0.05 were considered significant. Subsequently, bioinformatics analysis, including GO and KEGG pathway analysis, was performed to identify differentially expressed proteins (18).
Experimental verification of tissue repair proteins in fetal rats with artificial cleft lip wounds
The mRNA levels of the differentially expressed molecules were assessed using reverse transcription-quantitative (RT-q) PCR, as previously described (19). Differentially expressed levels of proteins were detected by label-free quantification PRM as previously described (17).
Statistical analysis
GraphPad Prism 8.0 (GraphPad Software, Inc.) and SPSS 22.0 (IBM Corp.) were used to perform calculations and carry out statistical analysis. Student's t-test was used to compare differences between two groups. The experimental data from each group were analyzed for congruence of variance before the t-test were applied. The FDR values were within 0.01 in the comparisons. Mixed ANOVA followed by Sidak's post hoc test was used to analyze the differences between multiple groups. P<0.05 was considered to indicate a statistically significant difference.
Results
Gross observation
All fetal rats were observed before delivery. The nasolabial cleft was first observed before surgery and images were captured to facilitate the observation of changes in the fetal rats from the GA16.5 and GA18.5 groups. We observed the same area again 72 h post-surgery to identify differences. The cuneiform tissue of the upper-left lip was removed by microsurgery to create a cleft lip wound. The changes in the fetal rats were observed macroscopically. In the GA16.5 group, the upper-left cleft lip wound completely healed 72 h after surgery (i.e., GA19.5) and the continuity of the upper lip tissue was restored. Only a slight depression was observed in the surgical area. The upper-left lip tissue was nearly symmetrical with that of the right side. However, in the GA18.5 group, the cleft lip wound was not completely healed 72 h after surgery (i.e., GA21.5); a clear scar was observed in the surgical area, and the upper lip was asymmetrical on both sides due to wound contracture (Fig. 1).
Histological analysis
In the GA16.5 group 72 h after surgery, the tissue of the upper-left lip wound demonstrated complete regeneration when observed under the microscope (Figs. 2–4). The results of H&E staining demonstrated complete epithelialization of the upper-left lip, and the structure of new follicles was detected under the epidermis. Compared with the normal skin of the upper-right lip, a slight depression in the cleft part of the upper-left lip and thickening of the skin was noted, whereas inflammatory cell infiltration and neovascularization were not apparent (Fig. 2). Masson's Trichrome staining revealed collagen fibers under the epidermis, demonstrating a fine reticular and emerging follicular structure (Fig. 3). Immunohistochemical analysis indicated no obvious difference in the amount of type-I collagen in the upper-left cleft lip area and the rest of the upper lip (Fig. 4).
In the GA18.5 group, the position of the wound was easily identified by a distinct scar on the upper-left lip. H&E staining demonstrated that partial epithelialization occurred in the upper-left cleft lip area. Compared with the normal skin of the upper-right lip, the upper-left lip displayed a clear scar, new capillary formation around the wound and increased fibroblast proliferation and ECM volume, whereas structural components of hair follicles were not observed under the epidermis (Fig. 5). Masson's Trichrome staining demonstrated the absence of new follicular structure and the presence of dense collagen fibers under the epidermis (Fig. 6). Immunohistochemical analysis in the upper-left cleft lip wound demonstrated an increase in type-I collagen expression and fiber density, as well as a more compact structure and absence of adnexal skin (Fig. 7), compared with normal upper lip tissue.
Immunohistochemical analysis of cell proliferation markers was also carried out. Compared with GA16.5 fetal rats, the expression of Ki67 and CD31 slightly increased in the GA18.5 group following surgery. By contrast, the expression of CK10 decreased in the GA18.5 group, compared with the GA16.5 group (Fig. 8).
RT-qPCR analysis of inflammatory factors
The relative mRNA expression levels of the pro-inflammatory factors TNF-α, IL-10 and TGF-β were evaluated in the two groups of fetal rats. The mRNA levels of TNF-α and IL-10 were significantly higher in GA18.5 rats, compared with GA16.5 rats. Furthermore, the mRNA expression levels of TGF-β were significantly reduced in the GA18.5 group (Fig. 9).
Protein identification and differential protein screening
Compared with group 1, 57 differentially expressed proteins were identified in group 2, of which 37 were upregulated and 20 were downregulated. A comparison of groups 3 and 4 revealed 312 differentially expressed proteins, of which 171 were upregulated and 141 were downregulated. Lastly, compared with group 1,289 differentially expressed proteins were identified in group 3, of which 151 were upregulated and 138 were downregulated. Only 50 differentially expressed proteins and their multiple variations were upregulated or downregulated between all groups (Tables I–IV). The distribution of the differentially expressed proteins among the selected samples is presented as volcano plots (Figs. 10,11,12).
Bioinformatics analysis
Gene ontology (GO) enrichment analysis was performed on the differentially expressed proteins, and their properties were generally described as biological process (BP), molecular function (MF) or cellular component (CC). The first 10 GO enrichment results from each group are displayed in Fig. 13. The results demonstrated that 73, 542 and 376 differentially expressed proteins were significantly enriched between groups 1 and 2, 3 and 4 and 3 and 1, respectively. The results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified the possible pathway related to the differentially expressed proteins between groups (Fig. 14).
In addition, the interaction network of the differentially expressed proteins that regulate wound repair were analyzed. Examples of the interaction networks of the differentially expressed proteins involved in wound repair are as follows: i) Smad4, Tgf1i1, Ptpn6 and Hmox1 in group 1 and 2; ii) S100a9, Fgg, Anxa1, Fgb, Plg and S100a8 in group 3 and 4; and iii) CD36, S100a9, S100a8, Cd9Fgg, Anxa1, Fgb, Plg and S100a8 in group 3 and 1 (Fig. 15).
RT-qPCR analysis of possible target protein in cleft lip repair
RNA was extracted from tissue samples with TRIzol® reagent and the quality was checked using gel electrophoresis. Relative mRNA levels were analyzed using RT-qPCR (Fig. 16). The relative mRNA expression levels of Smad4 were significantly higher in group 2, compared with group 1 (P<0.05). Moreover, the relative mRNA expression levels of Fabp5 were significantly lower in groups 4 and 1, compared with group 3 (P<0.05). Additionally, the relative mRNA expression levels of S100a4 were significantly lower in group 4, compared with group 3 (P<0.05). S100a8 and S100a9 were significantly higher in group 3, compared with in groups 1 and 4 (P<0.05).
Immunofluorescence results
Immunofluorescence staining of Smad4, Fabp5, S100a4, S100a8 and S100a9 was performed on samples from both the GA16.5 and GA18.5 groups 72 h post-surgery. The expression levels of all five proteins increased in GA18.5 compared to GA16.5, and the differences were statistically significant (P<0.05; Fig. 17).
PRM analysis of differential protein expression
The differences in multiple variations of Smad4 expression were compared between groups 1 and 2. The panel reaction monitoring calculated this difference as 0.557, indicating downregulation in group 1 compared with in group 2 (P=0.043) (Table II). In contrast, no statistically significant differences were observed between groups 3 and 4. The difference in multiple variations of Fabp5 between groups 3 and 4 was calculated as 2.91, indicating upregulation in group 3 compared with in group 4 (P=0.024) (Table III). Additionally, the expression levels of Fabp5 were upregulated (P=0.01) in group 3 compared with in group 1; however, the difference between the variations present in groups 1 and 2 was not statistically significant. The difference in the multiple variations of S100a4 and S100a8 between groups 3 and 4 was calculated as 2.897 and 92.828, respectively, indicating an upregulation of the expression levels of both proteins in group 3 (P=0.001 and P=0.002, respectively) (Table III). Furthermore, the difference in the multiple variations of S100a8 between groups 3 and 1 was 25.933, which indicates upregulation in group 3 (P=0.002) (Table IV). However, the differences were not statistically significant between groups 1 and 2. The difference in the multiple variations of S100a9 was 30.191 and 19.538 between groups 3 and 4 and groups 3 and 1, respectively, suggesting upregulation in group 3 (P=0.0004 and P=0.001, respectively) (Tables III and IV). In contrast, the difference in the multiple variations of S100a9 between groups 1 and 2 was not statistically significant (Tables II–IV).
Discussion
In recent decades, various animal models of congenital cleft lip have been successfully established through surgical induction (4,20). It has been suggested that intrauterine cleft lip repair can effectively improve this defect and reduce the impact of scars on normal facial development after birth. Thus, it also provides a new way for the effective repair of congenital cleft lips. In the present study, pregnant SD rats were used to establish a fetal rat model of cleft lip wound at two time points, GA16.5 and GA18.5. The different pregnancy models were induced by using two different repair methods of a cleft lip wound of the fetus (21,22). The exact gestational age is particularly important for the results of the repair of cleft lip in the fetal rats. Thus, the use of a rat model provides an added advantage in that the exact time of conception can be replicated, thereby minimizing differences between groups.
The present findings confirmed the hypothesis that fetal rat defects can be regenerated during early pregnancy without scar formation (23). It was also demonstrated that fetal rat defects could not be completely regenerated in late pregnancy and resulted in scarring (24). Furthermore, the expression of pro-inflammatory factors was different between the two groups. However, these observations were only made at one time point (72 h) after constructing cleft lip wounds in fetal rats. Future studies are needed to examine samples collected at different time points following surgery. Another shortcoming of this study entails the lack of comparison between the cleft lip wound repairs of fetal rats at different ages, such as the fetus in the early stages of pregnancy, or in newborn and/or adult rats. Label-free quantitative proteomics were used to examine proteins that play important roles in the postoperative repair process of fetal cleft lip. Protein expression was examined in four groups of samples. In addition, bioinformatics analysis was conducted to identify potential biological markers, providing a theoretical reference and methodological basis for the examination of relevant mechanisms underlying fetal intrauterine scar repair. However, further studies are required to determine whether any one protein or several proteins, plays a key role in wound healing.
Smad4 belongs to the family of Smad proteins and is a common mediator in the signal transduction processes of the TGF-β family (25). TGF-β expression can lead to fibroblast proliferation and ECM deposition (26,27). The present findings indicated that the mRNA and protein expression levels of Smad4 were downregulated in the scar-free repair group.
Furthermore, the mRNA and protein expression levels of Fabp5 were upregulated in the scar formation group. Therefore, it may be hypothesized that Fabp5 could be involved in the fibrosis of the fetal cleft lip wound, which may be mediated by the TGF-β signaling pathway (28–30).
S100a4 is a member of the S100 calcium-binding protein family, and its expression is associated with various non-neoplastic diseases, such as chronic obstructive pulmonary disease and cardiac hypertrophy (31–35). The present study demonstrated that the mRNA and protein expression levels of S100a4 were upregulated in the scar formation group, which may be associated with scar repair of fetal rat cleft lip wounds.
S100a8 is also a member of the S100 calcium-binding protein family (36–42). mRNA and protein expression levels of S100a8 were significantly upregulated in the scar repair group in the present study, indicating a potential role for S100a8 in the process of fetal cleft lip wound healing.
Current reports frequently associate S100a9, a member of the calcium-binding protein family S100, with infectious diseases, immune diseases and tumors, such as non-small cell lung adenocarcinoma (43–45). mRNA and protein expression levels of S100a9 were significantly upregulated in the scar formation group. Therefore, we speculated that S100a9 may play an important role in the process of fetal wound healing. However, whether the reduced expression levels of Fabp5, S100a4, S100a8 and S100a9 in the third trimester of pregnancy would reduce or worsen scar formation remains unclear. Further functional testing and regulatory studies are required to confirm the role of these five differentially expressed proteins in fetal wound repair.
The cleft lip is a very common congenital condition that often leaves life-long scarring. The present study identified five differentially expressed proteins, namely Smad4, Fabp5, S100a4, S100a8 and S100a9, that may be potential biomarkers of the scarless repair process in fetal rat cleft lip wounds. These findings may facilitate the discovery of new clinical targets for the prevention and treatment of scars. However, the role of these proteins in fetal wound repair and potential underlying mechanisms require further examination.
Acknowledgements
Not applicable.
Funding
The present study was supported by a grant from the National Science and Basic Resources Survey Special Foundation, China (grant no. 2017FY101204), the Technology Innovation Guide Program of Hunan Province, China (grant no. 2017SK50124), the Science and Technology Major Project of Hunan Province, China (grant no. 2019SK1010) and the Science and Technology Major Project of Hunan Province, China (grant no. 2019SK1015).
Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Authors' contributions
YY, FH and JZho conceived and designed research. YY and FH performed animal experiments and staining. YY, HL, PJ, FH, JY, KG, SH and JZha performed PCR and label-free quantification PRM. YY, FH, JC, JY, ZC, AW and JZha analyzed data. YY and FH prepared figures. YY drafted the manuscript. FH and JZho edited and revised the manuscript. YY, FH, HL AW, PJ and JZho approved the final version of the manuscript. FH and JZho confirmed the authenticity of all of the raw data. All authors read and approved the final manuscript.
Ethics approval and consent to participate
This study followed the regulations stipulated by the People's Republic of China regarding the Management of Experimental Animals and was approved by The Animal Experiment Management and Medical Ethics Sub-committee of The Third Xiangya Hospital of Central South University, Hunan, China (approval no. 2014-S168).
Patient consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
References
|
VanKoevering KK, Morrison RJ, Prabhu SP, Torres MF, Mychaliska GB, Treadwell MC, Hollister SJ and Green GE: Antenatal three-dimensional printing of aberrant facial anatomy. Pediatrics. 136:e1382–e1385. 2015. View Article : Google Scholar : PubMed/NCBI | |
|
Burrington JD: Wound healing in the fetal lamb. J Pediatr Surg. 6:523–528. 1971. View Article : Google Scholar : PubMed/NCBI | |
|
Beanes SR, Hu FY, Soo C, Dang CM, Urata M, Ting K, Atkinson JB, Benhaim P, Hedrick MH and Lorenz HP: Confocal microscopic analysis of scarless repair in the fetal rat: Defining the transition. Plast Reconstr Surg. 109:160–170. 2002. View Article : Google Scholar : PubMed/NCBI | |
|
Walmsley GG, Hu MS, Hong WX, Maan ZN, Lorenz HP and Longaker MT: A mouse fetal skin model of scarless wound repair. J Vis Exp. 16:522972015. | |
|
Dang CM, Beanes SR, Lee H, Zhang X, Soo C and Ting K: Scarless fetal wounds are associated with an increased matrix metalloproteinase-to-tissue-derived inhibitor of metalloproteinase ratio. Plast Reconstr Surg. 111:2273–2285. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Longaker MT, Whitby DJ, Adzick NS, Crombleholme TM, Langer JC, Duncan BW, Bradley SM, Stern R, Ferguson MW and Harrison MR: Studies in fetal wound healing, VI. Second and early third trimester fetal wounds demonstrate rapid collagen deposition without scar formation. J Pediatr Surg. 25:63–69. 1990. View Article : Google Scholar : PubMed/NCBI | |
|
Lorenz HP, Whitby DJ, Longaker MT and Adzick NS: Fetal wound healing. The ontogeny of scar formation in the non-human primate. Ann Surg. 217:391–396. 1993. View Article : Google Scholar : PubMed/NCBI | |
|
Cass D, Bullard KM, Sylvester KG, Yang EY, Longaker MT and Adzick NS: Wound size and gestational age modulate scar formation in fetal wound repair. J Pediatr Surg. 32:411–415. 1997. View Article : Google Scholar : PubMed/NCBI | |
|
Longaker MT and Adzick NS: The biology of fetal wound healing: A review. Plast Reconstr Surg. 87:788–798. 1991. View Article : Google Scholar : PubMed/NCBI | |
|
Armstrong JR and Ferguson MW: Ontogeny of the skin and the transition from scar-free to scarring phenotype during wound healing in the pouch young of a marsupial, Monodelphis domestica. Dev Biol. 169:242–260. 1995. View Article : Google Scholar : PubMed/NCBI | |
|
Stern M, Dodson TB, Longaker MT, Lorenz HP, Harrison MR and Kaban LB: Fetal cleft lip repair in lambs: Histologic characteristics of the healing wound. Int J Oral Maxillofac Surg. 22:371–374. 1993. View Article : Google Scholar : PubMed/NCBI | |
|
Longaker MT, Dodson TB and Kaban LB: A rabbit model for fetal cleft lip repair. J Oral Maxillofac Surg. 48:714–719. 1990. View Article : Google Scholar : PubMed/NCBI | |
|
Oberg KC, Evans ML, Nguyen T, Peckham NH, Kirsch WM and Hardesty RA: Intrauterine repair of surgically created defects in mice (lip incision model) with a microclip: Preamble to endoscopic intrauterine surgery. Cleft Palate Craniofac J. 32:129–137. 1995. View Article : Google Scholar : PubMed/NCBI | |
|
Hu F, Yan Y, Wang CW, Liu Y, Wang JJ, Zhou F, Zeng QH, Zhou X, Chen J, Wang AJ and Zhou JD: Article effect and mechanism of ganoderma lucidum polysaccharides on human fibroblasts and skin wound healing in mice. Chin J Integr Med. 25:203–209. 2019. View Article : Google Scholar : PubMed/NCBI | |
|
Xue YN, Yan Y, Chen ZZ, Chen J, Tang FJ, Xie HQ, Tang SJ, Cao K, Zhou X, Wang AJ and Zhou JD: LncRNA TUG1 regulates FGF1 to enhance endothelial differentiation of adipose-derived stem cells by sponging miR-143. J Cell Biochem. 120:19087–19097. 2019. View Article : Google Scholar : PubMed/NCBI | |
|
Guo H, Chen T, Liang Z, Fan L, Shen Y and Zhou D: iTRAQ and PRM-based comparative proteomic profiling in gills of white shrimp Litopenaeus vannamei under copper stress. Chemosphere. 263:1282702021. View Article : Google Scholar : PubMed/NCBI | |
|
Chen PS, Li YP and Ni HF: Morphology and evaluation of renal fibrosis. Adv Exp Med Biol. 1165:17–36. 2019. View Article : Google Scholar : PubMed/NCBI | |
|
Luo Y, Yan Y, Zhang S and Li Z: Computational approach to investigating key GO terms and KEGG pathways associated with CNV. Biomed Res Int. 2018:84068572018. View Article : Google Scholar : PubMed/NCBI | |
|
Dong X, Landford WN, Hart J, Risolino M, Kaymakcalan O, Jin J, Toyoda Y, Ferretti E, Selleri L and Spector JA: Toward microsurgical correction of cleft lip ex utero through restoration of craniofacial developmental programs. Plast Reconstr Surg. 140:75–85. 2017. View Article : Google Scholar : PubMed/NCBI | |
|
Stelnicki EJ, Lee S, Hoffman W, Lopoo J, Foster R, Harrison MR and Longaker MT: A long-term, controlled-outcome analysis of in utero versus neonatal cleft lip repair using an ovine model. Plast Reconstr Surg. 104:607–615. 1999. View Article : Google Scholar : PubMed/NCBI | |
|
Harling TR, Stelnicki EJ, Hedrick MH and Longaker MT: In utero models of craniofacial surgery. World J Surg. 27:108–116. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Mast BA, Haynes JH, Krummel TM, Diegelmann RF and Cohen IK: In vivo degradation of fetal wound hyaluronic acid results in increased fibroplasia, collagen deposition, and neovascularization. Plast Reconstr Surg. 89:503–509. 1992. View Article : Google Scholar : PubMed/NCBI | |
|
Frantz FW, Diegelmann RF, Mast BA and Cohen IK: Biology of fetal wound healing: Collagen biosynthesis during dermal repair. J Pediatr Surg. 27:945–949. 1992. View Article : Google Scholar : PubMed/NCBI | |
|
Wilgus TA: Regenerative healing in fetal skin: A review of the literature. Ostomy Wound Manage. 53:16–33. 2007.PubMed/NCBI | |
|
Hu HH, Chen DQ, Wang YN, Feng YL, Cao G, Vaziri ND and Zhao YY: New insights into TGF-beta/Smad signaling in tissue fibrosis. Chem Biol Interac. 292:76–83. 2018. View Article : Google Scholar : PubMed/NCBI | |
|
Honardoust D, Ding J, Varkey M, Shankowsky HA and Tredget EE: Deep dermal fibroblasts refractory to migration and decorin-induced apoptosis contribute to hypertrophic scarring. J Burn Care Res. 33:668–677. 2012. View Article : Google Scholar : PubMed/NCBI | |
|
Wu C, Jiang J, Boye A, Jiang Y and Yang Y: Compound Astragalus and Salvia miltiorrhiza extract suppresses rabbits' hypertrophic scar by modulating the TGF-β/Smad signal. Dermatology. 229:363–368. 2014. View Article : Google Scholar : PubMed/NCBI | |
|
Furuhashi M, Ogura M, Matsumoto M, Yuda S, Muranaka A, Kawamukai M, Omori A, Tanaka M, Moniwa N, Ohnishi H, et al: Serum FABP5 concentration is a potential biomarker for residual risk of atherosclerosis in relation to cholesterol efflux from macrophages. Sci Rep. 7:2172017. View Article : Google Scholar : PubMed/NCBI | |
|
Yeung DC, Wang Y, Xu A, Cheung SC, Wat NM, Fong DY, Fong CH, Chau MT, Sham PC and Lam KS: Epidermal fatty-acid-binding protein: A new circulating biomarker associated with cardio-metabolic risk factors and carotid atherosclerosis. Eur Heart J. 29:2156–2163. 2008. View Article : Google Scholar : PubMed/NCBI | |
|
Song J, Zhang H, Wang Z, Xu W, Zhong L, Cao J, Yang J, Tian Y, Yu D, Ji J, et al: The role of FABP5 in radiation-induced human skin fibrosis. Radiat Res. 189:177–186. 2018. View Article : Google Scholar : PubMed/NCBI | |
|
Fei F, Qu J, Li C, Wang X, Li Y and Zhang S: Role of metastasis-induced protein S100A4 in human non-tumor pathophysiologies. Cell Biosci. 7:642017. View Article : Google Scholar : PubMed/NCBI | |
|
Grotterød I, Maelandsmo GM and Boye K: Signal transduction mechanisms involved in S100A4-induced activation of the transcription factor NF-kappaB. BMC Cancer. 10:2412010. View Article : Google Scholar | |
|
Schneider M, Kostin S, Strøm CC, Aplin M, Lyngbaek S, Theilade J, Grigorian M, Andersen CB, Lukanidin E, Lerche Hansen J and Sheikh SP: S100A4 is upregulated in injured myocardium and promotes growth and survival of cardiac myocytes. Cardiovasc Res. 75:40–50. 2007. View Article : Google Scholar : PubMed/NCBI | |
|
Tomcik M, Palumbo-Zerr K, Zerr P, Avouac J, Dees C, Sumova B, Distler A, Beyer C, Cerezo LA, Becvar R, et al: S100A4 amplifies TGF-β-induced fibroblast activation in systemic sclerosis. Ann Rheum Dis. 74:1748–1755. 2015. View Article : Google Scholar : PubMed/NCBI | |
|
Zhao YX, Ho CK, Xie Y, Chen YH, Li HZ, Zhang GY and Li QF: Calcimycin suppresses S100A4 expression and inhibits the stimulatory effect of transforming growth factor β1 on Keloid fibroblasts. Ann Plast Surg. 81:163–169. 2018. View Article : Google Scholar : PubMed/NCBI | |
|
Donato R: Intracellular and extracellular roles of S100 proteins. Microsc Res Tech. 60:540–551. 2003. View Article : Google Scholar : PubMed/NCBI | |
|
Lin H, Andersen GR and Yatime L: Crystal structure of human S100A8 in complex with zinc and calcium. BMC Struct Biol. 16:82016. View Article : Google Scholar : PubMed/NCBI | |
|
Bouzidi F and Doussiere J: Binding of arachidonic acid to myeloid-related proteins (S100A8/A9) enhances phagocytic NADPH oxidase activation. Biochem Biophys Res Commun. 325:1060–1065. 2004. View Article : Google Scholar : PubMed/NCBI | |
|
Gebhardt C, Németh J, Angel P and Hess J: S100A8 and S100A9 in inflammation and cancer. Biochem Pharmacol. 72:1622–1631. 2006. View Article : Google Scholar : PubMed/NCBI | |
|
Basso D, Bozzato D, Padoan A, Moz S, Zambon CF, Fogar P, Greco E, Scorzeto M, Simonato F, Navaglia F, et al: Inflammation and pancreatic cancer: Molecular and functional interactions between S100A8, S100A9, NT-S100A8 and TGFβ1. Cell Commun Signal. 26:12–20. 2014.PubMed/NCBI | |
|
Shabani F, Farasat A, Mahdavi M and Gheibi N: Calprotectin (S100A8/S100A9): A key protein between inflammation and cancer. Inflamm Res. 67:801–812. 2018. View Article : Google Scholar : PubMed/NCBI | |
|
Yaundong L, Dongyan W, Lijun H and Zhibo X: Effects of downregulation of S100A8 protein expression on cell cycle and apoptosis of fibroblasts derived from hypertrophic scars. Aesthet Surg J. 34:160–167. 2014. View Article : Google Scholar : PubMed/NCBI | |
|
Hessian PA, Edgeworth J and Hogg N: MRP-8 and MRP-14, two abundant Ca (2+)-binding proteins of neutrophils and monocytes. J Leukoc Biol. 53:197–204. 1993. View Article : Google Scholar : PubMed/NCBI | |
|
Markowitz J and Carson WE III: Review of S100A9 biology and its role in cancer. Biochim Biophys Acta. 1835:100–109. 2013.PubMed/NCBI | |
|
Zhong A, Xu W, Zhao J, Xie P, Jia S, Sun J, Galiano RD, Mustoe TA and Hong SJ: S100A8 and S100A9 are induced by decreased hydration in the epidermis and promote fibroblast activation and fibrosis in the dermis. Am J Pathol. 186:109–122. 2016. View Article : Google Scholar : PubMed/NCBI |
