The effects of HA-TCP scaffold complexes loaded with periodontal membrane stem cells on periodontal tissue regeneration

doi:10.3877/cma.j.issn.2095-1221.2025.02.004

Abstract

Abstract:

Objective

To investigate the effects of hydroxyapatite tricalcium phosphate（HA-TCP） scaffold complexes and periodontal ligament stem cells on periodontal tissue regeneration.

Methods

Healthy premolars from 15 children aged 16-18 who required corrective treatment were collected.The human periodontal ligament stem cells （hPDLSCs）， were isolated and identified.The self-replication ability of hPDLSCs was detected by using clone formation experiments.Flow cytometry was used to detect the expression of surface antigens CD90， CD44， CD34， and CD45 on hPDLSCs.Alizarin red and red oil O staining were used to detect the osteogenic and adipogenic abilities of hPDLSCs.Alkaline phosphatase （ALP）staining was used to detect alkaline phosphatase activity.hPDLSCs and HA-TCP complexes were prepared.Rat alveolar bone defect models were established.Thirty rats were divide into three groups randomly： NC group （no implantation），HA- TCP group （only HA-TCP implantation）， and hPDLSCs/HA-TCP group （hPDLSCs/HA- TCP composite implantation）.Pathological changes in alveolar bone tissues were detected by HE staining， and the expression of ALP， osteocalcin （OCN）， and osteopontin （OPN） proteins in alveolar bone tissue were detected by Western blot.

Results

Crystal violet staining shows that hPDLSCs can form clone colonies， and the clone colony status was observed under the microscope， indicating that hPDLSCs have self replication ability.The flow cytometry results showed high levels of mesenchymal stem cell markers CD90 and CD44 on the surface of hPDLSCs， while hematopoietic stem cell markers CD45 and CD34 were expressed at low levels， indicating that the extracted PDLSCs were mesenchymal stem cells.Alizarin red and red oil O staining revealed bone nodules， calcium mineralization nodules， and lipid droplet formation in hPDLSCs.The HE staining results showed that hPDLSCs/HA TCP promoted the formation of mature new bone， new blood vessels， and osteoblasts；ALP staining revealed an increase in the number of positive cells.Western blot results showed that compared with the NC group， the expression of ALP （0.63 ± 0.08， 1.21 ± 0.13 vs 0.32 ± 0.05）， OCN（0.61 ± 0.08， 1.10 ± 0.12 vs 0.23 ± 0.04）， and OPN protein （0.53 ± 0.07， 0.82 ± 0.10 vs 0.20 ± 0.04）in the alveolar bone tissue of the HA-TCP group and hPDLSCs/HA-TCP group were increased， and the expression of ALP， OCN， and OPN proteins in the alveolar bone tissue of the hPDLSCs/HA- TCP group was higher than that of the HA-TCP group （P ＜ 0.05）.

Conclusion

HA- TCP/PDLSCs complex can promote new bone formation， repairing alveolar bone loss and promoting bone tissue regeneration.

Key words: HA-TCP, Periodontal ligament stem cells, Periodontal tissue regeneration, Osteogenic differentiation

Lifei Liao, Pengcheng Liao, Sasa Shi, Ruizhao Ma, Xinjun Qu. The effects of HA-TCP scaffold complexes loaded with periodontal membrane stem cells on periodontal tissue regeneration[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2025, 15(02): 93-99.

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Figures/Tables 7

References 24

1	罗敬一，程波.基于转录组学探讨氧化应激相关基因在牙周炎中的作用机制［J］.临床口腔医学杂志， 2023， 39（3）：147-152.
2	Fan R， Zhou Y， Chen X， et al.Porphyromonas gingivalis outer membrane vesicles promote apoptosis via msRNA-Regulated DNA methylation in periodontitis［J］.Microbiol Spectr， 2023，11（1）：e0328822.doi： 10.1128/spectrum.03288-22.
3	刘丹.牙周炎与全身系统性疾病相关性研究［J］.每周文摘·养老周刊， 2023，（3）：51-53.
4	陈子君，王以琴，李杰，等.口腔微生物所致牙周炎在冠心病发病中的机制研究进展［J］.中国动脉硬化杂志， 2023， 31（10）：909-915.
5	Perić Kačarević Ž， Rider P， Alkildani S， et al.An introduction to bone tissue engineering［J］.Int J Artif Organs， 2020， 43（2）：69-86.
6	Tavelli L， McGuire MK， Zucchelli G， et al.Extracellular matrix-based scaffolding technologies for periodontal and peri-implant soft tissue regeneration［J］.J Periodontol， 2020， 91（1）：17-25.
7	Ding T， Kang W， Li J， et al.An in situ tissue engineering scaffold with growth factors combining angiogenesis and osteoimmunomodulatory functions for advanced periodontal bone regeneration［J］.J Nanobiotechnology， 2021， 19（1）：247.doi： 10.1186/s12951-021-00992-4.
8	Zhang Y， Luo W， Zheng L， et al.Efficient bone regeneration of BMP9-stimulated human periodontal ligament stem cells （hPDLSCs） in decellularized bone matrix （DBM） constructs to model maxillofacial intrabony defect repair［J］.Stem Cell Res Ther， 2022， 13（1）：535.doi：10.1186/s13287-022-03221-3.
9	谢冰，杨振宇，郭世梁.CircBARD1 靶向miR-495-3p/TLR4 调控LPS 诱导的人牙周膜细胞增殖、凋亡和炎症［J］.临床口腔医学杂志，2023， 39（3）：138-143.
10	Lima JR， Soares PBF， Pinotti FE， et al.Comparison of the osseointegration of implants placed in areas grafted with HA/TCP and native bone［J］.Microsc Res Tech， 2022， 85（8）：2776-2783.
11	Helaehil JV， Lourenço CB， Huang B， et al.In vivo investigation of polymer-ceramic PCL/HA and PCL/β-TCP 3D composite scaffolds and electrical stimulation for bone regeneration［J］.Polymers （Basel）， 2021，14（1）：65.doi： 10.3390/polym14010065.
12	Jepsen S， Suvan J， Deschner J.The association of periodontal diseases with metabolic syndrome and obesity［J］.Periodontol 2000， 2020，83（1）：125-153.
13	Abdulkareem AA， Al-Taweel FB， Al-Sharqi AJB， et al.Current concepts in the pathogenesis of periodontitis： from symbiosis to dysbiosis［J］.J Oral Microbiol， 2023， 15（1）：2197779.doi：10.1080/20002297.2023.2197779.
14	Eftekhari A， Maleki Dizaj S， Sharifi S， et al.The use of nanomaterials in tissue engineering for cartilage regeneration； current approaches and future perspectives［J］.Int J Mol Sci， 2020， 21（2）：536.doi： 10.3390/ijms21020536.
15	Park KM， Cheong J， Pang NS， et al.Medication-related osteonecrosis of the jaw using periodontitis-induced rat before tooth extraction［J］.BMC Oral Health， 2023， 23（1）：561.doi： 10.1186/s12903-023-03200-x.
16	Tavelli L， McGuire MK， Zucchelli G， et al.Biologics-based regenerative technologies for periodontal soft tissue engineering［J］.J Periodontol， 2020， 91（2）：147-154.
17	Zhang Z， Deng M， Hao M， et al.Periodontal ligament stem cells in the periodontitis niche： inseparable interactions and mechanisms［J］.J Leukoc Biol， 2021， 110（3）：565-576.
18	Seo BM， Miura M， Gronthos S， et al.Investigation of multipotent postnatal stem cells from human periodontal ligament［J/OL］.Lancet，2004， 364 （9429）：149-155.
19	Liu J， Li Q， Liu S， et al.Periodontal ligament stem cells in the periodontitis microenvironment are sensitive to static mechanical strain［J/OL］.Stem Cells Int， 2017， 2017：1380851.doi：10.1155/2017/1380851.
20	He Y， Jian CX， Zhang HY， et al.Hypoxia enhances periodontal ligament stem cell proliferation via the MAPK signaling pathway［J/ OL］.Genet Mol Res， 2016， 15（4）.doi： 10.4238/gmr15048965.
21	闫伟，杨莉，凌梅，等.自体骨髓富集间质干细胞与羟基磷灰石磷酸三钙材料复合在脊柱融合中的应用［J］.中国组织工程研究，2016， 20（8）：1075-1081.
22	王德元，白峰，高文魁，等.复合血管内皮细胞的不同孔径磷酸钙陶瓷体内血管化的比较研究［J］.现代生物医学进展， 2014， 14（20）：3815-3818， 3901.
23	陈思奇，先德彬，徐荣胜，等.羟基磷灰石-磷酸三钙支架复合骨髓间充质干细胞和人脐静脉内皮细胞对大鼠颅骨缺损修复早期成血管的影响［J］.中国组织工程研究， 2021， 25（22）：3458-3465.
24	Yang JX， Xie P， Li YS， et al.Osteoclast-derived miR-23a-5p-containing exosomes inhibit osteogenic differentiation by regulating Runx2［J］.Cell Signal， 2020， 70：109504.doi： 10.1016/j.cellsig.2019.109504.

[1]	Xin Chen, Xiaochen Zhang, Wen Qin, Zuolin Jin. Overexpression of methyltransferase-like 3 repairs the osteogenic ability of periodontal mesenchymal stem cells in patients with periodontitis [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2023, 17(01): 15-25.
[2]	Chenglin Liu, Wenxing Xun, Haizhen Yang, Sumeng Fan, Yubo Liu, Hongmei Zhang. Necrostatin-1 promotes the proliferation and osteogenic differentiation of periodontal ligament stem cells in high-glucose environment [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2022, 16(03): 160-167.
[3]	Weiyang Chen, Jun Tian, Xi Wei. The role of silicon ion in the field of bone regeneration [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2021, 15(06): 375-381.
[4]	Runze Li, Jianhan Ren, Delan Huang, Haotian Luo, Chen Zhou, Weicai Wang. Research progress and prospects of long non-coding RNA regulating osteogenic differentiation [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2020, 14(05): 271-279.
[5]	Jing Zhang, Chenchen Li, Jian Meng. The effect of interleukin-1β on the osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2020, 14(02): 88-94.
[6]	Hong Zhang, Yu Yao, Jiadong Sun, Xiaonan Yu, Zhiguang Zhang. NELL-1 regulates the P1 promoter of RUNX2 to promote osteogenic differentiation [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2017, 11(04): 197-203.
[7]	Yanjiao Li, Lei Liang, Fang Jin, Zhiwei Wang. Effects of ginkgolide B on proliferation and osteogenic differentiation of human perimembrane stem cells by regulating miR-24-3p [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(04): 229-235.
[8]	Yang Yang, Cheng Wang, Wentu Zhou, Bing Zhou. Caveolae/Caveolin-1 regulates osteogenic differentiation of mouse BMSCs together with membrane cholesterol [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(03): 137-142.
[9]	Bing Zhou, Zheng Wang. DEPTOR promotes proliferation and osteogenic differentiation of human periodontal stem cells by interacting with ErbB2 [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(06): 358-364.
[10]	Yidong Liu, Yufeng Jia, Yan Wang, Yang Liu, Jinyue Ning. Upregulation of Wnt-1 gene expression by soybean isoflavones to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(05): 292-297.
[11]	Cong Zhu, Guofeng Huang, Huixiang Jiang, Benwen Wu, Jianbiao Lin, Weibin Lin, Mingming Gao, Zhenqi Ding. Intermittent axial compressive stress promotes adhesion, proliferation and osteogenic differentiation of seed cells in tissue engineered bone [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2018, 08(06): 334-342.
[12]	Weili Zhang, Min Luo, Jiang Peng, Bin Zhao. The BMP-2/Smads signaling pathway promote the osteogenic differentiation of mesenchymal stem cells in rat with bone metabolism imbalance [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2021, 07(02): 65-72.
[13]	Bo Wu, Ye Liu, Xu Ma, Chunsheng Zhi, Mingchang Du, Liangquan Zhai, Zhengbo Yang, Jiayuan Wang, Yihan Wang. Effect of 3D printed collagen / hydroxyapatite scaffold on osteogenic differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2020, 06(03): 123-127.
[14]	Ye Zhang, Xiumei Zhuang, Yue Zhang, Qin Wang, Xuezhen Peng. Hypoxia inhibits osteogenic differentiation of human periodontal ligament cells via p53 upregulation [J]. Chinese Journal of Clinicians(Electronic Edition), 2018, 12(09): 518-524.
[15]	Min Zhou, Donghui Wu, Yalin Wu, Jingwen Pang, Xiumei Zhuang. Hydrophilicity of titanium with micro/nanotopographical surface promotes proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells [J]. Chinese Journal of Clinicians(Electronic Edition), 2018, 12(02): 98-102.

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