研究指出，透明質酸可以促進角膜上皮細胞增生，並能幫助角膜傷口癒合。本研究欲建立一動物角膜輪部上皮傷口損傷模式，並進一步探討透明質酸分子量及濃度對於輪部上皮幹細胞之影響評估。實驗使用不同分子量 (4300D, 12kD, 230kD, 1500kD)及濃度 (5μg/cm2, 0.5μg/cm2)的透明質酸製備培養介面，培養輪部上皮幹細胞，經繼代培養三代觀察幹細胞標誌分子p63及分化分子K3表現，發現其中以分子量12kD，高濃度5μg/cm2培養介面下，p63表現維持一定高表現60~80%而K3表現量有下降的驅勢由80%~60%，並由胞落群聚(colony forming assay)實驗發現透明質酸12kD比其他分子量有較多的胞落出現表示其培養介面下可以使輪部上皮幹細胞維持幹細胞特性。在不同培養介面環境下進行MTT assay，觀察輪部上皮幹細胞的增生活性，發現分子量12kD介面相較控制組有較佳的增生能力。最後動物試驗，以透明質酸分子量12kD，治療輪部上皮傷口（15mm2）發現在第一天即有明顯傷口癒合之現象，組織切片結果顯示上皮呈現完整的複層化。此試驗結果顯示，透明質酸12kD具有促進輪部上皮幹細胞之增生，維持其幹細胞特性之潛能及促進輪部上皮傷口癒合之能力。

Hyalurnonan (HA) are reported to stimulate corneal epithelial cells to proliferation, and help corneal wound healing. The objective of this research was to set up a corneal limbal epithelial wound healing model, and evaluate the effects of different HA concentration and molecular weight on Limbal epithelial stem cells. This study used different molecular weights (4300D, 12kD, 230kD and 1500kD) and concentrations (5μg/cm2 and 0.5μg/cm2) cultured surfaces, to culture LSCs and passage 3 times, and record the expression of stem-cell associated marker, p63, and differential marker, K3. The data shows HA 12kD, 5μg/cm2 cultured surface making LSCs have high p63 expression 60%~80% and K3 expression decreasing 80% to 60%. Furthermore, the colony forming assay also show HA 12 kD culture surface occurs more colonies, it suggests that the culture surface can maintain LSCs keeping their stem cell property. In order to measuring LSCs proliferation activity on different culture surfaces, MTT assay was used and the data shows HA 12kD compared with control group has better effects. Finally, in the animal model, the treatment of HA 12kD on limbal epithelial wound (15 mm2), there is obvious wound healing on day 1, and after healing, the corneal and limbal epitheliums on HA 12 kD group have more intact stratified squamous epithelums. Overall, this study data shows HA 12kD can stimulate LSCs proliferation and maintain stemm cell property and accerate limbal epithelial wound healing.

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