組織再生 Tissue Regeneration

細胞 生長因子 培養 殖入 取得細胞

Growth Factors Biomaterials Tissue regeneration Cells 細胞 Growth Factors Biomaterials 生長因子 生物材料

生長因子 基因重組--PDGF 血小板---PDGF、VEGF、TGF-b1、 FGF 、EGF

血小板 a-顆粒 PDGF VEGF TGF-b1 FGF EGF

細胞 生長因子 小牛血清? 培養 殖入 取得細胞

Growth Factors Biomaterials Tissue regeneration Cells 細胞 Growth Factors Biomaterials 生長因子 生物材料 ?

生物醫學材料 無毒性、安全性 生物相容性、生物降解性 足夠之機械支撐力 促進組織再生 殺(抑)菌性 促進(保護)生長因子釋放(破壞)

生物醫學材料 膠原蛋白 玻尿酸 幾丁質、幾丁聚糖 PGL PGEL

利用真菌做為生物醫學材料 Application of fungal materials

Why fungal materials? Fungi provided wide range of products from food to drugs Cell wall composition Polysaccharides and chitin/chitosan (幾丁質、幾丁聚糖)

靈芝子實體細胞壁-SACCHACHITIN之應用

Fruiting bodies of Ganoderma ?

一般靈芝之應用 供健康食品用 但留下許多廢渣

Residue of extractions 廢渣如何利用? 主要為Cell wall組成

SACCHACHITIN之處理流程 1N NaOH H2O2 可製成膜狀

Chemical Analysis 化學分析 TLC GC GC/MS NMR

SACCHACHITIN 分析結果顯示主要成分如下 50-60% Polysaccharide 40-50% N-acetyl glucosamine

Is SACCHACHITIN safe and functional? 安全性及有效性 Safety Cytotoxicity test, Immunogenecity Animals Rat Guinea pig Canine Inhibition Enzyme and Microbes Human/Clinic trials

Cells 利用細胞測試毒性 Fibroblast (天竺鼠纖維母細胞) Keratinocyte (人類角質細胞) Corneal epidermis cell (角膜表皮細胞) 結果均顯示無毒性，並有促進增值之作用

安全 增生 天竺鼠 Fibroblasts

促進遷移 Fibroblasts Control SACCHACHITIN 遷移方向 BESCHITIN W B-Glucan

人類角質細胞也有相同結果 Growth curve of human keratinocyte in the addition 0.01% SACCHACHITIN suspension.

Fig.6. Growth of keratinocyte in suspension of SACCHACHITIN Control 6 days 6 days Fig.6. Growth of keratinocyte in suspension of SACCHACHITIN in evaluating of cytotoxicity.

無過敏反應及 抗體反應

有無促進傷口癒合之性質?

紗布 紗布 SACCHACHITIN 紗布 SACCHACHITIN 21 DAYS 7 DAYS

傷口面積變化 一種日本CHITINT商品 SACCHACHITIN

在Guinea pigs實驗也有類似結果

For pets 為寵物製造之 SACCHACHITIN 10%混合CMC軟膏

Day 1 Day 7 Day 30 Day 15 已使用在許多寵物 之案例 燙傷之流類犬

Day 13 三月齡混種雌犬4kg體重，疑似因不當保溫電燈泡散發出的輻射熱所引致之三度燙傷。 Day 30

Day 14 一歲齡混種雌犬，疑似因天冷長時間睡在電熱器前造成在右髖部及大腿焦痂皮膚潰瘍的病變，大小約為9 cm、9 cm、7 cm的三邊長的三角形。 Day 35

人體試驗 Patients Totally 47 patients were treated with SACCHACHITIN. All the patients suffered chronic skin ulcer for over 6 months with 3-10 cm wound in diameter. All the patients were treated with chemotherapy or other possible treatment, previously. 36 patients were with satisfactory healing. The other 11 patients were lost their contact.

66 years old male Infection after open chest operation for 7 months Treatment of SACCHACHITIN 2001/4/16 2001/4/28

Chronic ulcer of left limb for 1 year Male, 63 yr Old Misuse of steroid drug, Chronic ulcer of left limb for 1 year Daily treatment of SACCHACHITIN 2001/4/10 Evident granulation tissue regenerated 2001/5/7

Pressure sore-1 Male, 79 yr old 2002/6/27 2002/7/11 2002/9/19

Pressure sore-2 Female, 80 yr old 2002/8/1 2002/8/15 2002/9/15

What is the mechanism that SACCHACHITIN functioned as a wound healing accelerator? One of the properties of fungal cell wall is to binding lytic enzymes in environment as a defense.

Fungal cell wall configuration Chitin as network Polysaccharide as matrix Fungal cell wall configuration

To trap lytic enzymes (MMPs, proteases, lipase… from bacteria) ?

酵素環境 MMPs (Matrix Metallo-proteinases) MMP-2, MMP-9 the collagenases

Inflammation Early stage platelets aggregate blood coagulation Late stage Neutrophils, monocytes Macrophages---bacteria

Matrix metallo-proteases (MMPs) MMPs were produced by Neutrophils. MMPs hydrolyzed collagen piling as well as the growth factors. Association of MMPs and chronic ulcer has been reported.

Enzyme in vitro test supported that… 27kDa Binding effect of SACCHACHITIN on MMP-9

Quantitative analysis of MMP-9 protein levels in skin wound tissue