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Evaluation of the cell viability and in vivo biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats

  • Authors:
    • Sruthy Prathap
    • Rajesh Kashyap Shankar
    • Nebu George Thomas
    • Riaz Abdulla
    • Binsi Puthanpurakkalkizhakkethil Kamalamma
    • Suprith Surya
    • Prathap Mulackal Sreekanatan
  • View Affiliations / Copyright

    Affiliations: Department of Periodontology, Yenepoya Dental College, Yenepoya University, Mangalore, Karnataka 575018, India, Department of Periodontology, Pushpagiri College of Dental Sciences, Pushpagiri Institute of Medical Sciences and Research Centre, Thiruvalla, Kerala 689107, India, Department of Oral Pathology, Yenepoya University, Mangalore, Karnataka 575018, India, Fish Processing Division, Central Institute of Fisheries Technology, Willington Island, Cochin, Kerala 682029, India, Advanced Surgical Skill Enhancement Division, Yenepoya University, Mangalore, Karnataka 575018, India, Department of Conservative Dentistry and Endodontics, Yenepoya Dental College, Mangalore, Karnataka 575018, India
    Copyright: © Prathap et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 35
    |
    Published online on: March 10, 2026
       https://doi.org/10.3892/wasj.2026.450
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Abstract

The development of novel bone substitutes for the management of periodontal osseous defects requires assured biological function upon implantation. Chicken egg shells have been introduced for some time in reconstructive surgeries due to their mineral composition. Egg shells are among the most abundant byproducts of food processing waste and appear to have unique biological and chemical properties that support cell differentiation. Additives such as fish collagen and glycerine may enhance their efficacy by improving their regenerative properties, which mimic those of human bone, and increasing their handling characteristics. Biocompatibility and cell viability are two critical factors affecting the ability of any regenerative material to integrate into the body. The aim of the present study was to determine the cell viability and biocompatibility of three egg shell‑derived biomaterials, namely, egg shell‑derived hydroxyapatite (EHPA), EHPA modified with fish collagen EHPA/Coll, and EHPA/Coll with glycerine. Chicken egg shells were used to synthesise hydroxyapatite via chemical precipitation. The synthesised hydroxyapatite was modified with fish collagen and then with glycerol. All three materials were tested for their viability in mouse fibroblasts and for their in vivo biocompatibility in Wistar rats. The results revealed that eggshell‑derived hydroxyapatite was highly biocompatible and did not exert any cytotoxic effects. Its modification using fish collagen and glycerine also resulted in derivatives that were biocompatible and noncytotoxic. On the whole, the present study demonstrates that all three materials tested, which were eggshell derivatives, were highly biocompatible and promoted cell growth.
View Figures

Figure 1

Phase contrast images of the cells
treated with the indicated samples at x100 magnification. (A)
Control, (B) eggshell-derived hydroxyapatite, (C) composite graft
(EHPA/Coll), (D) composite graft with glycerine (EHPA/Coll/Gly).
EHPA, egg shell-derived hydroxyapatite; Coll, collagen; Gly,
glycerine.

Figure 2

Skin reactions following the placement
of materials. (A) Material placed in subcutaneous pouches; (B) 7th
day post-operative image (no notable skin reactions such as colour
changes, enema, rashes, or pus discharge were observed).

Figure 3

Histological representation of the
tissues from the implantation sites after 14 days. (A) Tissue from
untreated control sites, (B) tissue from sites treated with the
EHPA graft, (C) tissue from sites treated with the composite graft
(EHPA/Coll), (D) tissue from sites treated with the composite graft
with solvent (EHPA/Coll/Gly). Stereomicroscopy images (at x10
magnification) of the tissues collected from the animals, which
included the rat epithelium and connective tissue. The arrows
depict the inflammatory cells. EHPA, egg shell-derived
hydroxyapatite; Coll, collagen; Gly, glycerine.

Figure 4

Comparison of the mean inflammatory
scores between the groups. The Kruskall-Wallis test revealed no
significant difference between the groups (P>0.9999), although
slightly higher inflammatory scores were noted in the EHPA group
compared to the empty control (P=0.0137). *P<0.05
denotes a significant difference.

Figure 5

Correlation of the inflammatory scores
associated with different materials within the same animal.
Spearman's correlation revealed no correlation between the mean
inflammatory scores produced by different materials within the same
animal.
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Copy and paste a formatted citation
Spandidos Publications style
Prathap S, Shankar RK, Thomas NG, Abdulla R, Kamalamma BP, Surya S and Sreekanatan PM: Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats. World Acad Sci J 8: 35, 2026.
APA
Prathap, S., Shankar, R.K., Thomas, N.G., Abdulla, R., Kamalamma, B.P., Surya, S., & Sreekanatan, P.M. (2026). Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats. World Academy of Sciences Journal, 8, 35. https://doi.org/10.3892/wasj.2026.450
MLA
Prathap, S., Shankar, R. K., Thomas, N. G., Abdulla, R., Kamalamma, B. P., Surya, S., Sreekanatan, P. M."Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats". World Academy of Sciences Journal 8.3 (2026): 35.
Chicago
Prathap, S., Shankar, R. K., Thomas, N. G., Abdulla, R., Kamalamma, B. P., Surya, S., Sreekanatan, P. M."Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats". World Academy of Sciences Journal 8, no. 3 (2026): 35. https://doi.org/10.3892/wasj.2026.450
Copy and paste a formatted citation
x
Spandidos Publications style
Prathap S, Shankar RK, Thomas NG, Abdulla R, Kamalamma BP, Surya S and Sreekanatan PM: Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats. World Acad Sci J 8: 35, 2026.
APA
Prathap, S., Shankar, R.K., Thomas, N.G., Abdulla, R., Kamalamma, B.P., Surya, S., & Sreekanatan, P.M. (2026). Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats. World Academy of Sciences Journal, 8, 35. https://doi.org/10.3892/wasj.2026.450
MLA
Prathap, S., Shankar, R. K., Thomas, N. G., Abdulla, R., Kamalamma, B. P., Surya, S., Sreekanatan, P. M."Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats". World Academy of Sciences Journal 8.3 (2026): 35.
Chicago
Prathap, S., Shankar, R. K., Thomas, N. G., Abdulla, R., Kamalamma, B. P., Surya, S., Sreekanatan, P. M."Evaluation of the cell viability and <em>in vivo</em> biocompatibility of three potential regenerative biomaterials derived from eggshells in Wistar rats". World Academy of Sciences Journal 8, no. 3 (2026): 35. https://doi.org/10.3892/wasj.2026.450
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