| 摘要: | 背景：鈦植入材料在各種醫學治療中應用最為廣泛。在口腔保健中，植牙治療因其鄰牙的保存和良好的效果而成為修復缺牙的主要選擇。另一方面，黏膜周圍炎和感染相關的後果是目前植入治療對長期成功的有害影響。此外，快速骨整合是在短時間內交付最終假體的主要組成部分，因為傳統假體的負荷需要六個月以上。根據加速成骨細胞活性和抑制感染相關併發症的目的，採用不同的方法對植體表面進行修飾。鍶（Sr）是一種生物活性、非放射性材料，具有與鈣相似的生物效應，將其摻入生物醫學材料中可以增強骨整合。在微米級粗糙鈦表面上進行 Sr 塗層的奈米級改質在骨癒合過程和早期植體穩定性方面顯示出有希望的結果。 Sr可以增加成骨細胞的增殖並降低破骨細胞的活性。然而，對鈦表面摻鍶塗層的抗菌活性的研究有限。近年來，植體周圍黏膜炎和植體周圍炎在臨床實踐中變得更加常見。細菌的初始定植和早期感染是植牙失敗的主要原因。因此，尋求預防植體周圍區域微生物感染的策略非常重要。然而，抗生素可導致抗藥性細菌誘導和骨長入障礙。雙相磷酸鈣（BCP）是醫療應用中最常用的生物陶瓷之一，特別是在骨缺損、牙植體和牙周治療。 BCP 也具有與骨礦物相相似的成分，並廣泛影響骨誘導過程。 BCP 被認為是一種可生物降解材料，也可用作藥物輸送系統。由於溶膠-凝膠塗覆法是最有前途的製程之一，簡單，成本低，既可以小規模生產，也可以工業化大規模生產，因此採用溶膠-凝膠法將Sr-BCP摻入到Sr -BCP上。
研究目的：由於 Sr 和 BCP 都對骨整合產生正面影響，因此認為 Sr 和 BCP 的組合對成骨活性具有協同作用並增強骨整合。
材料與方法：牙科4級Ti圓盤（直徑15mm，厚度1mm）用SiC紙P240通過研磨機研磨。打磨後，使用臨床牙科噴砂機（TS）對鈦表面進行噴砂處理。將TS試片放入預先配製好的混合酸溶液（H2SO4+HCl）中作用24小時，並對材料（TSS）表面進行酸蝕處理。將 TSS 材料浸入 5M NaOH 溶液中 24 小時 (TSSA)。使用溶膠-凝膠方法，將圓盤塗上三種不同Sr 濃度（0.005M、0.01M 和0.02M）的Sr 摻入BCP，每組進一步細分為兩種不同的藻酸鹽(Alg) 濃度（1%和 3%）。然後放入烤箱，60℃烤12小時。分析物理性質，例如光學影像、SEM 和 EDS 分析。包括2個對照組和6個Sr-BCP塗層組在內的8組透過XRD和XPS分析、FTIR、潤濕性測試和粗糙度分析進一步分析了它們的物理和表面性能。透過使用 AlamarBlue 測定、鹼性磷酸? (ALP) 活性測定和茜素紅 S 染色 (ARS 測定) 測定來分析細胞活性。
結果：掃描電子顯微鏡分析發現，噴砂後鈦（Ti）的光滑表面變成了不規則的凹谷，部分刮痕區域明顯加深。隨後進行酸蝕，在低倍鏡下出現凹坑和裂縫。鹼處理後，凹坑間的隔膜變薄，呈現蜘蛛網狀結構。溶膠-凝膠塗層呈現結節狀外觀。在接觸角測量中，鹼處理後親水性增加。然而，親水性會根據溶膠-凝膠塗層後海藻酸鹽的百分比而變化。在用 Sr-CaP 化合物塗覆後，以 XRD 分析對三種 TiO2（銳鈦礦、金紅石和板鈦礦）、標準 HA 和 β-TCP 進行分析。在XPS分析中，化學成分和元素變得更高，顯示溶膠-凝膠塗層會影響表面化學結構。由於鹼處理後進行溶膠-凝膠塗層後可產生奈米級的 3D 表面，因此表面粗糙度變得更低。細胞增殖結果表明，改質 3D Ti 樣本和低藻酸鹽塗層 Sr-CaP 樣本的細胞生長有所改善。低 Alg % 的溶膠-凝膠塗層在細胞分化和細胞礦化分析方面均取得了良好的結果。
結論：在本研究的參數範圍內，低藻酸鹽百分比 (1%) 和較低鍶摩爾濃度 (0.01M) 的 Sr-CaP 塗層顯示出最佳的表面構型和細胞反應。此外，這些樣品的親水性顯示出良好的結果。溶膠-凝膠塗層後的 3D 結構表面的構建顯示出更好的結果，顯示了化學成分、親水性和表面元素，從而帶來更好的生物相容性和細胞活性。
BACKGROUND: Titanium implant materials are most widely used in a variety of medical treatments. In oral healthcare, dental implant treatment becomes the main choice for the replacement of missing teeth due to the preservation of adjacent teeth and exquisite outcomes. On the other hand, peri-mucositis and infection-related consequences are current deleterious effects of implant treatment for long-term success. Moreover, rapid osseointegration is the main component to deliver the final prostheses in a short time since the conventional prostheses loading takes more than six months. According to the purpose of accelerating the osteogenic cell activity and inhibiting infection-related complications, the implant surface is modified in different methods. Strontium (Sr) is a bioactive, nonradioactive material with and similar biological effect as calcium which can enhance osseointegration when it is incorporated into biomedical materials. Modifications at the nanoscale level with Sr coating on the micro rough titanium surface showed promising results in the bone healing process and early implant stability. Sr can increase the proliferation of osteoblastic cells and decrease osteoclast cell activity. However, there is limited study of the antimicrobial activities of Sr-incorporated coating on titanium surfaces. Recently, peri-implant mucositis and peri-implantitis become more common in clinical practice. Initial colonization and early infection of bacteria are major reasons for dental implant failure. Thus, it is important to seek strategies to prevent microbial infections in the peri-implant area. However, antibiotics can lead to resistant-bacterial induction and bone ingrowth disturbance. Biphasic calcium phosphate (BCP) is one of the most commonly used bioceramics in medical applications, especially in bone defects, dental implants, and periodontal therapy. BCP also has a similar composition to the mineral phase of bone and extensively influences the osteoinductive process. BCP is considered a biodegradable material and also acts as a drug delivery system. Since the sol-gel coating method is one of the most promising processes, simple, low cost, and can be obtained both in small-scale and industrial large-scale production, the sol-gel method was used to incorporate Sr-BCP on the nanoscale titanium surface to promote early osseointegration of titanium implants.
OBJECTIVE OF THE STUDY: Since both Sr and BCP influence osseointegration positively, the combination of Sr and BCP is assumed to have a synergistic effect on osteogenic activity and enhance osseointegration.
MATERIALS AND METHODS: Dental grade 4 Ti discs (15mm diameter and 1mm thickness) were ground with SiC paper P240 by a grinder. After grinding, Ti surfaces are sandblasted by using a clinical dental sandblasting machine (TS). TS test pieces are put in the mixed acid solution (H2SO4 + HCl) that has been prepared in advance and act for 24 hours to carry out the acid etching treatment on the surface of the material (TSS). TSS materials are dipped in 5M NaOH solution for 24 hours (TSSA). The discs are coated with Sr incorporated BCP in three different Sr concentrations (0.005M, 0.01M, and 0.02M) by using the Sol-Gel method, further sub-divided into two different alginate (Alg) concentrations for each group (1% and 3%). Then, put it in the oven at 60’C for 12 hours. Physical properties were analyzed, such as an optical image, SEM, and EDS analysis. 8 groups including the 2 control groups and 6 Sr-BCP coated groups further analyzed their physical and surface properties by using XRD and XPS analysis, FTIR, wettability test, and roughness analysis. The cell activities are analyzed by using AlamarBlue assay, Alkaline Phosphatase (ALP) Activity Assay, and Alizarin Red S Staining (ARS Assay) assay.
RESULTS: In SEM analysis, the smooth surface of titanium (Ti) changed into an irregular valley and some scratch areas prominently deepened after sandblasting. Followed by acid etching, pits, and fissures appeared in low magnification. After alkali treatment, the septums between pits became thinner and showed a spider web structure. Sol-gel coating displayed a nodules-like appearance. In contact angle measurement, properties of hydrophilicity increased after alkali treatment. However, the hydrophilicity changed according to the alginate percentages after the sol-gel coating. Three TiO2 (anatase, rutile, and brookite), standard HA, and β-TCP are analyzed in XRD analysis after coating with Sr-CaP compound. In XPS analysis, the chemical composition and the elements become higher showing that the sol-gel coating affects the surface chemical structure. The surface roughness becomes lower because of the 3D surface production at the nanoscale level after alkali treatment followed by sol-gel coating. The cell proliferation results show an improvement in cell growth of the modified 3D Ti sample and the Sr-CaP coated in low alginate coating. The sol-gel coating in low Alg % describes favorable results in both cell differentiation and cell mineralization analysis.
CONCLUSION: Within the parameters of the present investigation, Sr-CaP coating in low alginate percentage (1%) with lower strontium molarity (0.01M) displays the best surface configuration and cell response among others. Moreover, the hydrophilicity of these samples shows good results. The construction of the 3D structure surface followed by sol-gel coating shows better results showing the chemical compositions, hydrophilicity, and surface elements which lead to better biocompatibility and cellular activity. |
