β-glucan in wound healingβ-glucan in wound healing

Ri Ri 謝博帆謝博帆

Wound healingWound healing

β-glucanβ-glucan a fiber-type complex sugar (polysaccharide) derived a fiber-type complex sugar (polysaccharide) derived

from the cell wall of plants, fungi, and certain bacteriafrom the cell wall of plants, fungi, and certain bacteria

Physiology of β-glucanPhysiology 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 levelLowering cholesterol level Modifying neoplastic diseaseModifying neoplastic disease

Effect of β-glucan on wound Effect of β-glucan on wound healing healing

Activation of fibroblastActivation 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 strengthdeposition, increase tensile strength Stimulation of macrophageStimulation of macrophage

Applications of β-glucanApplications of β-glucan

I. Effectiveness of beta-glucan collagen for treatment of I. Effectiveness of beta-glucan collagen for treatment of partial-thickness burns in childrenpartial-thickness burns in children

II. Bio-artificial skin composed of gelatin and (1→3), II. Bio-artificial skin composed of gelatin and (1→3),

(1→6)-β-glucan(1→6)-β-glucan

I. β-glucan collagen for treatment of I. β-glucan collagen for treatment of partial-thickness burns of childrenpartial-thickness burns of children

BGC = beta-glucan collagen matrixBGC = beta-glucan collagen matrix

BGC as a temporary wound BGC as a temporary wound dressing, intended for management dressing, intended for management of partial-thickness burns, donor of partial-thickness burns, donor sites, and shallow woundssites, and shallow wounds

DiscussionDiscussion Outcome of burn injuries was not affected adversely by Outcome of burn injuries was not affected adversely by

the choice of BGC or standard treatment with silver the choice of BGC or standard treatment with silver sulfadiazinesulfadiazine

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 - provide a semiocclusive wound covering, decrease evaporative water and heat lossevaporative 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 II. Bio-artificial skin composed of gelatin and (1→3), (1→6)-β-glucan gelatin and (1→3), (1→6)-β-glucan

Dermal substitute:Dermal substitute:

function as a guide for cells moving into the repair area, function as a guide for cells moving into the repair area, serve as a scaffold, and help synthesize extracellular serve as a scaffold, and help synthesize extracellular matrix (ECM)matrix (ECM)

Porous gelatin/β-glucan sponge containing fibroblastPorous gelatin/β-glucan sponge containing fibroblast

(gelatin: denatured type of collagen, activating macrophages, (gelatin: denatured type of collagen, activating macrophages, and having hemostatic effect)and having hemostatic effect)

Material and methodsMaterial and methods

Sponge cultured with fibroblast: artificial dermisSponge cultured with fibroblast: artificial dermis Dermal equivalent cultured with keratinocyte: artificial Dermal equivalent cultured with keratinocyte: artificial

epidermisepidermis 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)Result (1)

Cellular attachment and proliferationCellular attachment and proliferation

Gel 100 & Gg91

Result (2)Result (2) Morphology of scaffoldsMorphology of scaffolds

Gel 100 Gg 91

Gg 73 Gg 55

Result (3)Result (3)

Distribution of fibroblast for 2 weeksDistribution of fibroblast for 2 weeks

Result (4)Result (4)

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

Result (5)Result (5)

Keratinocytes and fibroblasts cultured in Gg91 spongeKeratinocytes and fibroblasts cultured in Gg91 sponge

Result (6)Result (6)

H&E stain of mouse skin woundH&E stain of mouse skin wound

Gel 100

Gg 55

Gel 100 without fibroblasts

Discussion (1) Discussion (1) Cell attachment and proliferation depend on the Cell attachment and proliferation depend on the contact contact

angleangle, the , the chargecharge on the material surface, on the material surface, functional functional groupsgroups, and the , and the surface texturesurface texture

- Moderate contact angle: - Moderate contact angle: controlled by mixing the appropriate ratios of gelatin andβ-controlled by mixing the appropriate ratios of gelatin andβ-

glucan 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 binding with integrin receptor, promoting adhesion, regulating

cell growth, differentiationcell growth, differentiation

→ → Gel 100 & Gg91 are good for cell attachment and proliferationGel 100 & Gg91 are good for cell attachment and proliferation

Discussion (2)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 - form a dense surface (protection from evaporation from the wound, while having a detrimental effect on cell the wound, while having a detrimental effect on cell migration) → low concentration (0.7 wt%)migration) → low concentration (0.7 wt%)

Discussion (3)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 improve the re-epithelialization on the full-thickness defectdefect

Conclusion Conclusion

β-glucan promotes wound healing by activation of β-glucan promotes wound healing by activation of macrophages and fibroblastsmacrophages and fibroblasts

Bio-artificial skin containg β-glucan Bio-artificial skin containg β-glucan

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Normal human fibroblasts express pattern recognition receptors for fungal (1→3)-β-D-Normal human fibroblasts express pattern recognition receptors for fungal (1→3)-β-D-glucans. Infect Immunity 69:3933-3938, 2001glucans. Infect Immunity 69:3933-3938, 2001

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Lee SB, Jeon HW, Lee YW, Lee YM, Song KW, Park MH, Nam YS, Ahn HC Bio-artifical 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. skin composed of gelatin and (1→ 3),(1→6)-β-glucan . Biomaterials 24:2503-2511, 2003.

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