β-glucan in wound healing Ri 謝博帆 Ri 謝博帆

Wound healing

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

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

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

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

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

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

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

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

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

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

Result (3) Distribution of fibroblast for 2 weeks Distribution of fibroblast for 2 weeks

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

Result (5) Keratinocytes and fibroblasts cultured in Gg91 sponge Keratinocytes and fibroblasts cultured in Gg91 sponge

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

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

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

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

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

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: , 2001 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: , 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: , 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: , 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: , 2002a 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: , 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: , 2002b 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: , 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: , 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: , 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 (1) 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 (1)