The Effect of Human Amniotic Membrane, Epidermal Cells and Marrow Mesenchymal Stem Cells in Healing a Skin Defect

Chul Hong Kim; Sung Soo Kim; Sung Keun Shon; Dal Hee Kim; Chang Geun Song; Hyun Jun Kim

doi:10.4055/jkoa.2008.43.3.276

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Kim, Kim, Shon, Kim, Song, and Kim: The Effect of Human Amniotic Membrane, Epidermal Cells and Marrow Mesenchymal Stem Cells in Healing a Skin Defect

Original Article

Journal of the Korean Orthopaedic Association

Journal of the Korean Orthopaedic Association 2008; 43(3): 276-286.

Published online: 30 June 2008

DOI: https://doi.org/10.4055/jkoa.2008.43.3.276

The Effect of Human Amniotic Membrane, Epidermal Cells and Marrow Mesenchymal Stem Cells in Healing a Skin Defect

Chul Hong Kim, M.D., Sung Soo Kim, M.D., Sung Keun Shon, M.D., Dal Hee Kim, M.D., Chang Geun Song, M.D., Hyun Jun Kim, M.D.

Department of Orthopaedic Surgery, College of Medicine, Dong-A University, Busan, Korea.

Address reprint requests to Sung Soo Kim, M.D. Department of Orthopedic Surgery, College of Medicine, Dong-A University, 3-1, Dongdaesin-dong, Seo-gu, Busan 602-715, Korea. Tel: +82.51-240-2953, Fax: +82.51-254-6757, sskim2@dau.ac.kr

Abstract

Purpose

The purpose of this study was to investigate the transplantation results of human amniotic membrane (HAM), epidermal cells, or marrow mesenchymal stem cells (MSCs) in healing a skin defect.

Materials and Methods

Defects (full-thickness) in rabbits were treated with HAM alone (group A), HAM injected with cultivated epidermal cells (group B), HAM injected with cultivated MSCs (group C), or Vaseline gauze (group D). Tissue granulation, regeneration, re-epithelization and healing time were measured. Defects and healed area were calculated 2 weeks after surgery.

Results

The mean healing area was 67.5%, 81.7%, 83.2% and 49.5% in each group, with all treatment groups significantly different than group D (p<0.01), and groups B and C compared higher than group A (p<0.05). The healing time of groups A, B, and C was 5.7 to 6.4 days faster than that of group D (p<0.01). Histologic analysis showed that the new epidermis covered nearly the whole wound surface in group B and C, and contained granulated tissue with fibroblasts, capillaries, and collagen.

Conclusion

HAM grafts injected with cultivated epidermal cells or MSCs promoted healing of skin defects.

Keywords: Amniotic membrane, Epidermal cells, MSCs, Skin defect

Figures and Tables

Fig. 1

(A) Incision of a 3×3 cm size of full-thickness skin for collection of epidermal cells. (B) Aspiration and collection of 10 cc bone marrow mesenchymal stem cells in the rabbit femur.

Fig. 2

Photographs of cultivated epidermal cells and mesenchymal stem cells in vitro. (A) Epidermal cells (total cells: 6.8×10⁶, 5 cc). (B) Mesenchymal stem cells (total cells: 6.8×10⁶, 5 cc).

Fig. 3

A full-thickness skin defect of 2×2 cm in the rabbit.

Fig. 4

Graft procedures in each group. (A) Sutured graft of human amniotic membrane. (B) Injection of epidermal cells or mesenchymal stem cells under the human amniotic membrane. (C) Wound covered with Vaseline gauze.

Fig. 5

Comparison of HAM alone (A), HAM with epidermal cells (B), HAM with MSCs (C), and the Vaseline gauze (D) group 1 week post-graft (HAM, human amniotic membrane; MSCs, mesenchymal stem cells).

Fig. 6

Comparison of HAM alone (A), HAM with epidermal cells (B), HAM wih MSCs (C), and the vaseline gauze (D) group 2 weeks post-graft (HAM, human amniotic membrane; MSCs, mesenchymal stem cells).

Fig. 7

Histologic finding in group A. (A) Healed tissue shows a moderately recovered, relatively thin epidermis, but a completely healed skin appendage (×40). (B) Increased vascularity, little lymphocytic infiltration, and minimal fibrosis are noted in the dermis (×200).

Fig. 8

Histologic finding in group B. (A) Healed tissue shows a regenerated epidermis and recovered skin appendage (×40). (B) The dermis shows a few infiltrated inflammatory cells and mild fibrosis (×200).

Fig. 9

Histologic finding in group C. (A) Healed tissue shows a regenerated epidermis and recovered skin appendage (×40). (B) The dermis shows mild lymphoplasma cell infiltration and hemorrhaging; with abundant vasculature and minimal fibrosis are noted (×200).

Fig. 10

Histologic finding in group D. (A) Healed tissue shows an irregular and flattened ridge in the epidermis and the skin appendage is mostly absent (×40). (B) The dermis shows marked fibrosis with some lymphocyte infiltration and poor vasculature (×200).

Table 1

Mean Remaining Defect Area and Healing Area of Wounds at 2 Weeks Post-graft

Group D compared with group A, B and C. p<0.01, Group A compared with group B and C. p<0.05.

HAM, human amniotic membrane; MSCs, mesenchymal stem cells; Ep, epidermal cells; Va, Vaseline gauze.

Table 2

Mean Wound Healing Time

Group D compared with group A, B and C. p<0.01.

HAM, human amniotic membrane; MSCs, mesenchymal stem cells; Ep, epidermal cells; Va, Vaseline gauze.

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