本論文開發一包有PLGA微粒之鑲嵌式玻尿酸微針系統，可進行雙重骨鬆藥物之經皮傳輸，應用於骨質疏鬆症之長期治療。微針主體包覆親水性骨鬆藥物(alendronate sodium, ALN)以及PLGA微粒，PLGA微粒則包覆疏水性骨鬆藥物(estradiol, E2)，後端再結合快溶型玻尿酸組成的支撐軸，支撐軸能提供額外的長度以及足夠的機械強度，以抵抗皮膚之變形，使得微針完全鑲嵌，支撐軸快速溶解於皮膚中，持續緩釋ALN及E2。本研究製備的PLGA微米粒子具有高包覆效率(99.49 ± 1.5%)、高產率(99.68 ± 0.41%)以及粒徑分佈均一(1-20 μm)且無聚集的現象；由血中雌二醇濃度結果證實，本研究製備包有雌激素之PLGA微粒可於體內緩釋雌激素至少28天；微針能鑲嵌於皮膚中，深度約700 ~ 800 μm；將微針包覆model drug(coumarin、nile blue)後刺入老鼠背部皮膚，持續釋放親水染劑(Nile blue)長達14天，疏水染劑(coumarin)長達28天；在治療骨鬆方面，共分為5組，分別A(non-OVX)、B(OVX-PBS)、C(OVX-ALN)、D(OVX-MPs) and E(OVX-MNs).由生化標誌(biomarker) : CTX-I濃度方面可看出E組(2.11 ± 0.13 ng/ml)確實比C組(2.32 ± 0.12 ng/ml)、D組(2.77 ± 0.13 ng/ml)下降許多(P < 0.01)；微電腦斷層掃描儀(Micro-CT):骨密度(bone mineral density)方面可看出，E組(0.404 ± 0.026)確實比C組(0.302 ± 0.024)、D組(0.224 ± 0.026)上升許多(P < 0.01)，故由上述生化標誌(biomarker)、微電腦斷層掃描儀(Micro-CT)證實，微針組(協同治療)刺激成骨細胞及對破骨細胞的雙重抑制效果，使得微針組確實比單一給藥組(C、D組)具有更好的療效。本研究開發之包覆微米粒子之玻尿酸微針，確實可鑲嵌於皮膚內，進行長效且雙效型地釋放ALN及E2，對骨質疏鬆症做更有效的治療，並期許成為臨床上治療骨質疏鬆症的選擇之一。

This study develops an embeddable hyaluronic acid microneedle (HA MN) system containing PLGA microparticles (MPs) as a dual-drug transdermal delivery system for long-term treatment of osteoporosis. Alendronate sodium (ALN) and estradiol (E2)-loaded PLGA MPs are loaded in the HA MN that are connected with a dissolving HA supporting array. When inserted into the skin, the supporting array can provide an extended length to counteract skin deformation for complete insertion of MN and then quickly dissolve in the skin. The MN can be embedded within the skin for sustained transdermal delivery of ALN and E2. The prepared PLGA MPs has high drug entrapment efficiency of 99.49 ± 1.5%, high yield of 99.68 ± 0.41%, uniform particle size distribution (1-20 μm) and no aggregation between MPs. Serum E2 profiles show that the PLGA MNs allows prolonged release of E2 for at least 28 days. The MNs can pierce into porcine cadaver skin and rat skin at a depth of 700 ~ 800 μm. After being inserted into rat skin, the embedded MNs allowed sustained release of hydrophilic (Nile blue) and hydrophobic (coumarin) model drugs for up to 14 and 28 days, respectively. In anti-osteoporosis therapy test in ovariectomized (OVX) rat, we have five groups, namely A(non-OVX)、B(OVX-PBS)、C(OVX-ALN)、D(OVX-MPs) and E(OVX-MNs). Biomarker study shows the CCTX-I in E group (2.11 ± 0.13 ng/ml) was lower than C (2.32 ± 0.12 ng/ml)、D (2.77 ± 0.13 ng/ml) group (P < 0.01)； In Micro-CT study, bone mineral density (BMD) in E group (0.404 ± 0.026) was higher than C (0.302 ± 0.024)、D (0.224 ± 0.026) group (P < 0.01). These results demonstrated that the E group (dual-drug therapy) has a better therapy than the single drug-administrated group (C,D group) by stimulating osteoblasts and dual inhibition effect on osteoclasts. To sum up, the MNs loaded with PLGA-MPs have the ability to sustain delivery of dual-drug in the skin for long-term and better therapy of osteoporosis and we hope our MNs system can become an alternative way for long-term treatment of osteoporosis on clinical.

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