Interaction of Staphylococcus aureus with osteoblasts (Review)

  • Authors:
    • Sifeng Shi
    • Xianlong Zhang
  • View Affiliations

  • Published online on: December 20, 2011     https://doi.org/10.3892/etm.2011.423
  • Pages: 367-370
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Orthopedic infection is refractory to cure. Staphylococcus aureus (S. aureus) is the main causative pathogen responsible for orthopedic infection. S. aureus is capable of not only colonizing bone matrix, but also invading osteoblasts, which may play a significant role in the persistence and recurrence of osteomyelitis. Internalization requires the involvement of cytoskeletal elements, including actin microfilaments, microtubules and clathrin-coated pits. Microfilaments are most significant in the invasion process. S. aureus is capable of remaining alive in osteoblasts for a long period of time. Decreased sensitivity to antibiotics capable of penetrating host cells increases the difficulties of eradicating S. aureus. Osteoblasts, invaded by S. aureus, play a significant role in the initiation and maintenance of inflammatory immune responses. These osteoblasts recruit leukocytes and phagocytes to the site of inflammation via the expression of cytokines. Apoptosis is observed in osteoblasts invaded by S. aureus. Recruitment of osteoclasts and other immunocytes plays a crucial role in the resorption and destruction of bone.

Contents

Introduction

Invasion of osteoblasts by S. aureus

The fate and viability of S. aureus following invasion

The role of osteoblasts in the immune response

Apoptosis of osteoblasts following the internalization of S. aureus

Destruction of the bone due to inflammation and osteoclasts

Conclusion

Introduction

Orthopedic infection is a refractory disease that is associated with progression and recurrence (1). With increasing numbers of foreign bodies implanted in orthopedic surgery, such as prosthetic joints, implant-related infection poses a threat to patients (2). Deep infection following arthroplasty is severe and thorough debridement and removal of the foreign body should be carried out (3).

Staphylococcus aureus (S. aureus) is the main causative pathogen responsible for orthopedic infection (4). S. aureus is capable of colonizing the matrix of the bone. This infection is difficult to eradicate since bacterial biofilm formation and bacteria embedded in biofilms become resistant to antibiotics, leading to the persistence of osteomyelitis (5). However, S. aureus was shown to invade osteoblasts, which may play a significant role in the persistence and recurrence of osteomyelitis (68).

The main function of osteoblasts is to synthesize bone matrix and regulate the activity of osteoclasts (9). However, the ability of S. aureus to be internalized by host cells and the expression of cytokines reveal the significant role of osteoblasts in the immune response.

Invasion of osteoblasts by S. aureus

Although not considered to be a typical intracellular pathogen, S. aureus was found to be capable of invading osteoblasts and other host cells in vitro (2,4,1015). Internalization of bacteria by osteoblasts also occurs in vivo (16). In one experiment, S. aureus were injected subcutaneously under the skin of the scalp and the allantoic sac of 17-day-old chick embryos. Following 48 h, calvariae and tibiae were collected for transmission electron microscopy (TEM). S. aureus cells were found in approximately 14% of the calvarial osteoblasts following subcutaneous injection, and in 11% of calvarial and tibial osteoblasts following intra-allantoic injection (16). Internalization of S. aureus by osteoblasts have also been reported in a patient with recurrent, long-term osteomyelitis (17).

This phenomenon was considered to be another mechanism for the persistence and recurrence of chronic osteomyelitis. Bacteria and osteoblasts play a significant role in the invasion process. The attachment of S. aureus to osteoblasts is the first step of internalization and this attachment involves surface molecules of S. aureus. Fowler (18) proposed a model of attachment in which osteoblasts form a fibronectin bridge between surface-associated fibronectin-binding proteins of bacteria and host cell β1 integrins; this bridge then leads to the invasion of S. aureus. Staphylococcus epidermidis (S. epidermidis) was also shown to be capable of invading bone cells. However, unlike S. aureus, S. epidermidis was unable to gain entrance into bone cells through a fibronectin bridge between the integrin and a bacterial adhesin (19).

Different S. aureus strains have variable capacity in internalizing bacteria. This capacity is found to be correlated with σ B expressed by S. aureus (20). σ B, the only σ factor identified in S. aureus, is capable of prolonging the production of cell surface proteins, including fibronectin. σ B also plays a significant role in the regulation of virulence genes in S. aureus (21).

Internalization requires the involvement of cytoskeletal elements, including actin microfilaments, microtubules and clathrin-coated pits. Microfilaments are most significant in the invasion process (13). The process may be inhibited by monodansylcadaverine and cytochalasin D and, to some extent, by ouabain, monensin, colchicine and nocodazole (12). Notably, the invasion process does not require live bacteria (live and dead S. aureus are equally effective for invasion); however, live osteoblasts are required (22). The role of the osteoblast is positive in the process of invasion, and a ‘zipper-type’ mechanism has been proposed (23).

Calcium channels are also crucial in the invasion of osteoblasts, which are involved in the rearrangement of the cytoskeleton (24). The invasion of osteoblasts by S. aureus resulted in an increase in the phosphorylation of the extracellular signal-regulated protein kinases (ERK 1 and 2) (25). Activation of ERK 1 and 2 may result in the phosphorylation of numerous different substrates, including the transcription factors ATF-2, Elk-1 and c-Jun (26). ERK 1 and ERK 2 phosphorylation may also activate phospholipase A2, resulting in the production of leukotrienes (27). Leukotrienes may then open calcium channels on the host cell membrane (28).

The fate and viability of S. aureus following invasion

The fate of S. aureus following internalization by osteoblasts may correlate with the clinical manifestation of osteomyelitis. If S. aureus is capable of remaining alive in the intracellular environment, a thorough cure of osteomyelitis is difficult due to the difficulty of eradicating the bacteria in the osteoblast. Rifampin, chloramphenicol and clindamycin are the most active intracellular antibiotics. The majority of other antibiotics currently used are inactive intracellularly (e.g., lincomycin) or are incapable of penetrating cells (e.g., penicillins, cephalosporins and aminoglycosides) (29). Therefore, bacteria internalized by the osteoblast may be sequestered from the majority of antibiotics as well as the immune system.

Certain antibiotics are capable of eukaryotic cell penetration and have intracellular functions. However, similar to the bacteria embedded in the biofilms, the internalized bacteria may change their characteristics and decrease their sensitivity to antibiotics following the invasion of host cells and become situated in the intracellular environment. Investigators have used clindamycin and rifherampin to treat the S. aureus internalized in the osteoblast (30). The two antibiotics are capable of penetrating eukaryotic cells. Although the antibiotics may decrease or kill S. aureus internalized in the osteoblast at immediately following the invasion, the bacteria become less sensitive to the two antibiotics 12 h following the invasion (30). The observed structural changes were considered to be responsible for the change in sensitivity; however, metabolic change may also play a significant role. The change in sensitivity of S. aureus to antibiotics in the intracellular environment increases the difficulties of eradicating the pathogen.

Since S. aureus is capable of remaining alive in the osteoblast for a long period of time (4,22), the bacteria released from the dead osteoblast may induce the recurrence of osteomyelitis (6). A previous study also showed the lack of decreased viability of S. aureus in invading a second osteoblast in vitro (31).

The role of osteoblasts in the immune response

As is commonly known, the primary roles of osteoblasts are to synthesize the components of the bone matrix and to regulate osteoclasts, which are bone resorption cells (32). However, there is increasing understanding of the function of osteoblasts in the initiation and maintenance of the inflammatory immune response. Osteocytes may recruit leukocytes and phagocytes to the site of inflammation via the expression of cytokines.

Cultured mouse and human osteoblasts infected with S. aureus were found to express high levels of interleukin (IL-6 and IL-12p75) (33). IL-12 is capable of stimulating T lymphocytes and natural killer (NK) cells to secrete significant amounts of interferon (IFN), activating macrophages and T lymphocytes to augment a Th1 response(33). Although IL-12 is known for its ability to protect against intracellular pathogens, it may contribute to the process of organ-specific autoimmune diseases (34). Monocyte chemoattractant protein-1 (MCP 1), produced by osteoblasts, also has the ability to recruit macrophages and certain T lymphocytes to areas of inflammation (35,36). Osteoblasts may respond to bacterial infection by upregulating the expression of the chemokine CXCL10 (IP-10). IP-10 may then recruit T lymphocytes to the sites of bone infections (37). Dexamethasone, PGE(2) and T(h)2 cytokines are potential down-regulatory mediators of the chemokine (38).

Expression of NLRP3 in osteoblasts invaded by S. aureus was found in a study by McCall et al (39). The active NLRP3 inflammasome drives the innate immune response towards invading pathogens and cell damage, and regulates an adaptive immune response (40). The expression of NOD, a novel intracellular pattern recognition receptor, and Rip2 kinase, a critical downstream effector molecule for NOD signaling, was observed in osteoblasts invaded by S. aureus (41). NOD may regulate pro-inflammatory pathways in response to bacteria by inducing signalling pathways including nuclear factor κB (NF-κB) and mitogen-activated protein kinases (MAPKs) (42). However, certain authors believe it is the attachment, not invasion or secreted soluble factor(s) that activates NF-κB in human osteoblasts (43,44).

Apoptosis of osteoblasts following the internalization of S. aureus

Osteoblasts invaded by few bacteria were found to be able to remain alive and differentiate into osteocytes (16). However, apoptosis or programmed cell death was found in osteoblasts invaded by S. aureus, but the process may also be induced by pathogens. Tucker et al (45) used light microscopy to examine morphological changes in the osteoblasts following the internalization of S. aureus. Cell rounding was observed, and dark centers, due to condensation of chromatin, were noted. Apoptotic nuclei were also present.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is crucial in the process of cell apoptosis. Alexander et al (8) examined the ability of S. aureus to induce the production of TRAIL by osteoblasts. Results demonstrated that S. aureus was capable of inducing TRAIL expression by osteoblasts. A dose-dependent response was observed 30 min following exposure to bacteria. Attachment of S. aureus to osteoblasts is necessary for optimal TRAIL induction (46). Messenger RNA (mRNA) molecules encoding TRAIL receptors were also expressed by osteoblasts. The expression of NLRP3 in osteoblasts invaded by S. aureus also provides a potential mechanism of apoptotic cell death of the host cells (39). Osteoblast apoptosis results in decreased matrix deposition and destruction of the bone (8).

Destruction of the bone due to inflammation and osteoclasts

Normal bone remodeling requires the coordinated regulation of the genesis and activity of osteoblast and osteoclast lineages (47). Therefore, apoptosis of osteoblasts is not the only cause of bone destruction. Recruitment of osteoclasts and other immunocytes play a significant role in the resorption and destruction of bone. In S. aureus infection, bone resorption is caused by proteins rather than lipoteichoic acid or muramyl dipeptide. The surface-assosiated protein fraction may stimulate fibroblasts or monocytes to release osteolytic cytokines and chemokines (48), including MCP-1, colony-stimulating factors (CSFs) and interleukins. Dexamethasone, PGE(2) and T(h)2 cytokines have potential down-regulatory mediation of these chemokines (38).

Although MCP-1 is capable of recruiting macrophages and certain T lymphocytes to areas of inflammation (35,36), T lymphocytes and macrophages may also be responsible for bone loss and contribute to the development of the progressive inflammatory damage (35). IL-6, produced by osteoblasts invaded by S. aureus, may directly or indirectly modulate the activity of osteoclasts, resulting in the induction of osteoclast differentiation or osteoclast-mediated bone demineralization (49). IL-6 expressed in the osteoblasts may be suppressed by certain agents including epigallocatechin gallate, a constituent of tea, which plays a role in the suppression of inflammation and decrease of bone resorption (50). IL-12 also contributes to the process of organ-specific autoimmune diseases (34). CSFs have a profound effect on osteoclastogenesis, high levels of granulocyte-macrophage-CSF (GM-CSF) and G-CSF secretion by osteoblasts challenged by S. aureus, and may induce osteoclastogenesis, resulting in bone resorption (51).

Conclusion

The interaction of S. aureus with osteoblasts is initiated from the attachment of the bacteria to osteoblasts, followed by the internalization of S. aureus by osteoblasts. Signal transduction and cytokine expression play a vital role in the process of inflammatory damage and destruction of the bone.

S. aureus, internalized in the osteoblasts, may be sequestered from the majority of antibiotics and the immune system. However, the sensitivity change of the bacteria to the antibiotics capable of penetrating eukaryotic cells results in the failure of antibiotic therapy. Therefore, metabolic characteristics of S. aureus in the intracellular environment should be further studied to develop novel antibiotics to kill the bacteria in the osteoblasts. In addition, the internalization process should also be considered; if we are able to prohibit the invasion process and locate the bacteria outside the host cells, then eradication of these bacterias becomes simpler.

Osteoblasts challenged by S. aureus play a significant role in the initiation and maintenance of the immune response. The process is complicated and involves numerous cytokines and pathways that conduct the signal. Although recruitment of leukocytes and phagocytes may be helpful in killing the pathogens, inflammatory damage may accumulate. Apoptosis of osteoblasts induced by S. aureus and the increased activity of osteoclasts are responsible for bone resorption (39). Down-regulation of cytokines may inhibit the destruction of bone; however, down-regulation of cytokines also inhibits the immune response, which may be helpful in the process of eradicating bacteria (32). Therefore, it appears more promising and easier to inhibit the attachment and internalization of S. aureus by osteoblasts in treating osteomyelitis. Further investigation into the interaction of S. aureus with osteoblasts should therefore be performed in order to decrease the rate of orthopedic infection.

References

1. 

S ChiharaJ SegretiOsteomyelitisDis Mon56631201010.1016/j.disamonth.2009.07.001

2. 

N RaoBH ZiranBA LipskyTreating osteomyelitis: antibiotics and surgeryPlast Reconstr Surg127Suppl 1S177S187201110.1097/PRS.0b013e3182001f0f

3. 

G StocksHF JanssenInfection in patients after implantation of an orthopedic deviceASAIO J46S41S46200010.1097/00002480-200011000-0003611110293

4. 

LX WebbW WagnerD CarrollH TylerF ColdrenE MartinOsteomyelitis and intraosteoblastic Staphylococcus aureusJ Surg Orthop Adv1673782007

5. 

RA BradyGA O’MayJG LeidML PriorJW CostertonME ShirtliffResolution of Staphylococcus aureus biofilm infection using vaccination and antibiotic treatmentInfect Immun7917971803201121220484

6. 

C GarzoniWL KelleyStaphylococcus aureus: new evidence for intracellular persistenceTrends Microbiol175965200910.1016/j.tim.2008.11.005

7. 

S AhmedS MeghjiRJ WilliamsB HendersonJH BrockSP NairStaphylococcus aureus fibronectin binding proteins are essential for internalization by osteoblasts but do not account for differences in intracellular levels of bacteriaInfect Immun6928722877200110.1128/IAI.69.5.2872-2877.2001

8. 

EH AlexanderJL BentoFM HughesI MarriottMC HudsonKL BostStaphylococcus aureus and Salmonella enterica serovar dublin induce tumor necrosis factor-related apoptosis-inducing ligand expression by normal mouse and human osteoblastsInfect Immun6915811586200110.1128/IAI.69.3.1581-1586.2001

9. 

CV GayVR GilmanT SugiyamaPerspectives on osteoblast and osteoclast functionPoult Sci7910051008200010.1093/ps/79.7.100510901202

10. 

RA AlmeidaKR MatthewsE CifrianAJ GuidrySP OliverStaphylococcus aureus invasion of bovine mammary epithelial cellsJ Dairy Sci7910211026199610.3168/jds.S0022-0302(96)76454-8

11. 

B SinhaM HerrmannMechanism and consequences of invasion of endothelial cells by Staphylococcus aureusThromb Haemost94266277200516113815

12. 

JK EllingtonSS ReillyWK RampMS SmeltzerJF KellamMC HudsonMechanisms of Staphylococcus aureus invasion of cultured osteoblastsMicrob Pathog263173231999

13. 

M JevonC GuoB MaN MordanSP NairM HarrisB HendersonG BentleyS MeghjiMechanisms of internalization of Staphylococcus aureus by cultured human osteoblastsInfect Immun67267726811999

14. 

JA WrightSP NairInteraction of staphylococci with boneInt J Med Microbiol300193204201010.1016/j.ijmm.2009.10.00319889575

15. 

JK EllingtonM HarrisMC HudsonS VishinLX WebbR SherertzIntracellular Staphylococcus aureus and antibiotic resistance: implications for treatment of staphylococcal osteomyelitisJ Orthop Res2487932006

16. 

SS ReillyMC HudsonJF KellamWK RampIn vivo internalization of Staphylococcus aureus by embryonic chick osteoblastsBone2663702000

17. 

MJ BosseHE GruberWK RampInternalization of bacteria by osteoblasts in a patient with recurrent, long-term osteomyelitis. A case reportJ Bone Joint Surg Am8713431347200510.2106/JBJS.D.0264915930546

18. 

T FowlerER WannD JohS JohanssonTJ FosterM HöökCellular invasion by Staphylococcus aureus involves a fibronectin bridge between the bacterial fibronectin-binding MSCRAMMs and host cell beta1 integrinsEur J Cell Biol796726792000

19. 

H KhalilR WilliamsG StenbeckB HendersonS MeghjiS NairInvasion of bone cells by Staphylococcus epidermidisMicrobes Infect9460465200710.1016/j.micinf.2007.01.002

20. 

SP NairM BischoffMM SennB Berger-BachiThe sigma B regulon influences internalization of Staphylococcus aureus by osteoblastsInfect Immun7141674170200310.1128/IAI.71.7.4167-4170.200312819110

21. 

Q ZhangY FengQ ZhouQ LuoX ZhangL Qin[Contribution of sigma B to environmental stress tolerance in Listeria monocytogenes - a review]Wei Sheng Wu Xue Bao (In Chinese)49128212882009

22. 

MC HudsonWK RampNC NicholsonAS WilliamsMT NousiainenInternalization of Staphylococcus aureus by cultured osteoblastsMicrob Pathog194094191995

23. 

B SinhaM HerrmannMechanism and consequences of invasion of endothelial cells by Staphylococcus aureusThromb Haemost94266277200516113815

24. 

BB FinlayP CossartExploitation of mammalian host cell functions by bacterial pathogensScience276718725199710.1126/science.276.5313.7189115192

25. 

JK EllingtonA ElhofyKL BostMC HudsonInvolvement of mitogen-activated protein kinase pathways in Staphylococcus aureus invasion of normal osteoblastsInfect Immun6952355242200110.1128/IAI.69.9.5235-5242.200111500391

26. 

BJ PulvererJM KyriakisJ AvruchE NikolakakiJR WoodgettPhosphorylation of c-jun mediated by MAP kinasesNature353670674199110.1038/353670a01922387

27. 

CA RouzerT MatsumotoB SamuelssonSingle protein from human leukocytes possesses 5-lipoxygenase and leukotriene A4 synthase activitiesProc Natl Acad Sci USA83857861198610.1073/pnas.83.4.8573006030

28. 

J PaceMJ HaymanJE GalanSignal transduction and invasion of epithelial cells by S. typhimuriumCell72505514199310.1016/0092-8674(93)90070-78382566

29. 

CB WilsonRF JacobsAL SmithCellular antibiotic pharmacologySemin Perinatol62052131982

30. 

JK EllingtonM HarrisMC HudsonS VishinLX WebbR SherertzIntracellular Staphylococcus aureus and antibiotic resistance: implications for treatment of staphylococcal osteomyelitisJ Orthop Res2487932006

31. 

JK EllingtonM HarrisL WebbIntracellular Staphylococcus aureus. A mechanism for the indolence of osteomyelitisJ Bone Joint Surg Br859189212003

32. 

I MarriottOsteoblast responses to bacterial pathogens: a previously unappreciated role for bone-forming cells in host defense and disease progressionImmunol Res30291308200410.1385/IR:30:3:29115531771

33. 

KL BostWK RampNC NicholsonJL BentoI MarriottMC HudsonStaphylococcus aureus infection of mouse or human osteoblasts induces high levels of interleukin-6 and interleukin-12 productionJ Infect Dis18019121920199910.1086/315138

34. 

S TrembleauT GermannMK GatelyL AdoriniThe role of IL-12 in the induction of organ-specific autoimmune diseasesImmunol Today16383386199510.1016/0167-5699(95)80006-97546194

35. 

I MarriottDL GrayDM RatiOsteoblasts produce monocyte chemoattractant protein-1 in a murine model of Staphylococcus aureus osteomyelitis and infected human bone tissueBone37504512200510.1016/j.bone.2005.05.01116027056

36. 

D RossiA ZlotnikThe biology of chemokines and their receptorsAnnu Rev Immunol18217242200010.1146/annurev.immunol.18.1.21710837058

37. 

NA GasperCC PettyLW SchrumI MarriottKL BostBacterium-induced CXCL10 secretion by osteoblasts can be mediated in part through toll-like receptor 4Infect Immun7040754082200210.1128/IAI.70.8.4075-4082.200212117914

38. 

KM WrightJS FriedlandRegulation of chemokine gene expression and secretion in Staphylococcus aureus-infected osteoblastsMicrobes Infect6844852200410.1016/j.micinf.2004.04.00815374006

39. 

SH McCallM SahraeiAB YoungOsteoblasts express NLRP3, a nucleotide-binding domain and leucine-rich repeat region containing receptor implicated in bacterially induced cell deathJ Bone Miner Res233040200810.1359/jbmr.071002

40. 

C JinRA FlavellMolecular mechanism of NLRP3 inflammasome activationJ Clin Immunol30628631201010.1007/s10875-010-9440-320589420

41. 

I MarriottDM RatiSH McCallSL TranguchInduction of Nod1 and Nod2 intracellular pattern recognition receptors in murine osteoblasts following bacterial challengeInfect Immun7329672973200510.1128/IAI.73.5.2967-2973.2005

42. 

L Le BourhisS BenkoSE GirardinNod1 and Nod2 in innate immunity and human inflammatory disordersBiochem Soc Trans35147914842007

43. 

C GiulianiG NapolitanoI BucciV MontaniF Monaco[Nf-κB transcription factor: role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications]Clin Ter (In Italian)1522492532001

44. 

R NingX ZhangX GuoQ LiAttachment of Staphylococcus aureus is required for activation of nuclear factor κB in human osteoblastsActa Biochim Biophys Sin (Shanghai)428838922010

45. 

KA TuckerSS ReillyCS LeslieMC HudsonIntracellular Staphylococcus aureus induces apoptosis in mouse osteoblastsFEMS Microbiol Lett1861511562000

46. 

EH AlexanderFA RiveraI MarriottJ AnguitaKL BostMC HudsonStaphylococcus aureus - induced tumor necrosis factor - related apoptosis - inducing ligand expression mediates apoptosis and caspase-8 activation in infected osteoblastsBMC Microbiol35200310.1186/1471-2180-3-5

47. 

SP NairS MeghjiM WilsonK ReddiP WhiteB HendersonBacterially induced bone destruction: mechanisms and misconceptionsInfect Immun2371238019968698454

48. 

S NairY SongS MeghjiSurface-associated proteins from Staphylococcus aureus demonstrate potent bone resorbing activityJ Bone Miner Res107267341995

49. 

LC HofbauerAE HeufelderIntercellular chatter: osteoblasts, osteoclasts and interleukin 6Eur J Endocrinol134425426199610.1530/eje.0.13404258640290

50. 

I IshidaC KohdaY YanagawaH MiyaokaT ShimamuraEpigallocatechin gallate suppresses expression of receptor activator of NF-κB ligand (RANKL) in Staphylococcus aureus infection in osteoblast-like NRG cellsJ Med Microbiol56104210462007

51. 

KL BostJL BentoJK EllingtonI MarriottMC HudsonInduction of colony-stimulating factor expression following Staphylococcus or Salmonella interaction with mouse or human osteoblastsInfect Immun6850755083200010.1128/IAI.68.9.5075-5083.200010948128

Related Articles

Journal Cover

March 2012
Volume 3 Issue 3

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Shi S and Zhang X: Interaction of Staphylococcus aureus with osteoblasts (Review). Exp Ther Med 3: 367-370, 2012.
APA
Shi, S., & Zhang, X. (2012). Interaction of Staphylococcus aureus with osteoblasts (Review). Experimental and Therapeutic Medicine, 3, 367-370. https://doi.org/10.3892/etm.2011.423
MLA
Shi, S., Zhang, X."Interaction of Staphylococcus aureus with osteoblasts (Review)". Experimental and Therapeutic Medicine 3.3 (2012): 367-370.
Chicago
Shi, S., Zhang, X."Interaction of Staphylococcus aureus with osteoblasts (Review)". Experimental and Therapeutic Medicine 3, no. 3 (2012): 367-370. https://doi.org/10.3892/etm.2011.423