β-glucan in wound healing Ri 謝博帆

1. β-glucan in wound healing Ri 謝博帆

2. Wound healing

3. β-glucan • a fiber-type complex sugar (polysaccharide) derived from the cell wall of plants, fungi, and certain bacteria

4. Physiology of β-glucan • Immunomodulator: - activation of macrophage - phygocytosis of neutrophil - NK cell, complement system - activation of lymphocyte • Lowering cholesterol level • Modifying neoplastic disease

5. Effect of β-glucan on wound healing • Activation of fibroblast - transcription factor (AP-1, Sp1, NF-κB, NF-1) → production of PDGF-A&B, FGF-a&b, TGF-α&β, VEGF, NT-3, etc. → macrophage infiltration, reepithelialization, collagen deposition, increase tensile strength • Stimulation of macrophage

7. Applications of β-glucan I. Effectiveness of beta-glucan collagen for treatment of partial-thickness burns in children II. Bio-artificial skin composed of gelatin and (1→3), (1→6)-β-glucan

8. I. β-glucan collagen for treatment of partial-thickness burns of children • BGC = beta-glucan collagen matrix • BGC as a temporary wound dressing, intended for management of partial-thickness burns, donor sites, and shallow wounds

11. Discussion • Outcome of burn injuries was not affected adversely by the choice of BGC or standard treatment with silver sulfadiazine • Advantage of BGC: - lack of daily wound care (painful dressing) - provide a semiocclusive wound covering, decrease evaporative water and heat loss - effective barrier to bacterial contamination - allow improved physical and occupational therapy - lower financial expense

12. II. Bio-artificial skin composed of gelatin and (1→3), (1→6)-β-glucan • Dermal substitute: function as a guide for cells moving into the repair area, serve as a scaffold, and help synthesize extracellular matrix (ECM) • Porous gelatin/β-glucan sponge containing fibroblast (gelatin: denatured type of collagen, activating macrophages, and having hemostatic effect)

13. Material and methods • Sponge cultured with fibroblast: artificial dermis • Dermal equivalent cultured with keratinocyte: artificial epidermis • Animal test: - full-thickness wound, 1cm in diameter, on the back of mice - H&E stain of the wound tissue after 1 week

14. Result (1) • Cellular attachment and proliferation Gel 100 & Gg91

15. Result (2) • Morphology of scaffolds Gel 100 Gg 91 Gg 73 Gg 55

16. Result (3) • Distribution of fibroblast for 2 weeks

17. Result (4) • Masson’s trichrome staining for extracellular matrix (ECM) of Gg91 with fibroblasts

18. Result (5) • Keratinocytes and fibroblasts cultured in Gg91 sponge

19. Result (6) • H&E stain of mouse skin wound Gel 100 Gg 55 Gel 100 without fibroblasts

20. Discussion (1) • Cell attachment and proliferation depend on the contact angle, the charge on the material surface, functional groups, and the surface texture - Moderate contact angle: controlled by mixing the appropriate ratios of gelatin andβ-glucan - Positive charge of acidic residues of gelatin - Arginine-glycine-aspartic acid (RGD) group of gelatin: binding with integrin receptor, promoting adhesion, regulating cell growth, differentiation → Gel 100 & Gg91 are good for cell attachment and proliferation

21. Discussion (2) • β-glucan: - modification of hydrophilic state - modification of pore morphology - bioacitve wound healing • Gelatin: - promote wound healing - form a dense surface (protection from evaporation from the wound, while having a detrimental effect on cell migration) → low concentration (0.7 wt%)

22. Discussion (3) • Fibroblasts in dermal equivalent: - stimulate epidermal differentiation - release cytokines, accelerating the healing process - enhance the attachment of keratinocyte → improve the re-epithelialization on the full-thickness defect

23. Conclusion • β-glucan promotes wound healing by activation of macrophages and fibroblasts • Bio-artificial skin containg β-glucan

24. Reference • Kougias P, Wei D, Rice PJ, Ensley HE, Kalbfleisch J, Willams DL, Browder IW. Normal human fibroblasts express pattern recognition receptors for fungal (1→3)-β-D-glucans. Infect Immunity 69:3933-3938, 2001 • Delatte SJ, Evans J, Hebra A, Adamson W, Othersen HB, Tagge EP. Effectiveness of beta-glucan collagen for treatment of partial-thickness burns in children. J Pediatr Surg 36:113-118, 2001. • Wei D, Williams D, Browder W. Activation of AP-1 and SP1 correlates with eound growth factor gene expression in glucan-treated human fibroblasts. Int Immunopharmacology 2:1163-1172, 2002a • Wei D, Zhang L, Williams D, Browder W. Glucan stimulates human dermal fibroblast collagen biosythesis through a nuclear factor-1 depenent mechanism . Int Immunopharmacology 10:161-168, 2002b • Lee SB, Jeon HW, Lee YW, Lee YM, Song KW, Park MH, Nam YS, Ahn HC Bio-artifical skin composed of gelatin and (1→ 3),(1→6)-β-glucan . Biomaterials 24:2503-2511, 2003. • Pillai, R.; Redmond, M.; Röding, J. Anti-Wrinkle Therapy: Significant New Findings in the Non-Invasive Cosmetic Treatment of Skin Wrinkles with Beta-Glucan. Int. journal of cosmetic science, Volume 27, Number 5, October 2005, pp. 292-292(1)