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Prevention and treatment of peri-implant fibrosis by functionally inhibiting skeletal cells expressing the leptin receptor

Nature Biomedical Engineering volume 8, pages 1285–1307 (2024)Cite this article

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Abstract

The cellular and molecular mediators of peri-implant fibrosis—a most common reason for implant failure and for surgical revision after the replacement of a prosthetic joint—remain unclear. Here we show that peri-implant fibrotic tissue in mice and humans is largely composed of a specific population of skeletal cells expressing the leptin receptor (LEPR) and that these cells are necessary and sufficient to generate and maintain peri-implant fibrotic tissue. In a mouse model of tibial implantation and osseointegration that mimics partial knee arthroplasty, genetic ablation of LEPR+ cells prevented peri-implant fibrosis and the implantation of LEPR+ cells from peri-implant fibrotic tissue was sufficient to induce fibrosis in secondary hosts. Conditional deletion of the adhesion G-protein-coupled receptor F5 (ADGRF5) in LEPR+ cells attenuated peri-implant fibrosis while augmenting peri-implant bone formation, and ADGRF5 inhibition by the intra-articular or systemic administration of neutralizing anti-ADGRF5 in the mice prevented and reversed peri-implant fibrosis. Pharmaceutical agents that inhibit the ADGRF5 pathway in LEPR+ cells may be used to prevent and treat peri-implant fibrosis.

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Fig. 1: LEPR-expressing cells are present abundantly in both human and murine peri-implant fibrous tissue.
Fig. 2: Ablation of LEPR+ cells resulted in significant reduction of peri-implant fibrosis.
Fig. 3: Fibrous-integration surgery does not affect osteogenicity of Lin−LepR-tdTomato−6C3−CD90−CD105−CD200+.
Fig. 4: Engrafted Lin−LEPR-tdTomato+ cells from a fibrous-integrated donor are more inclined to form fibrous tissue than Lin–LEPR-tdTomato+ cells from a osseointegrated donor.
Fig. 5: ADGRF5 is expressed in abundance by LEPR+ cells in both human and murine peri-implant fibrotic tissue.
Fig. 6: Ablation of Adgrf5 in LEPR+ cells inhibits peri-implant fibrosis and enhances peri-implant bone formation in vivo.
Fig. 7: Ablation of Adgrf5 in LEPR+ resulted in decreased LEPR+ cell proliferation and increased osteogenic differentiation capability.
Fig. 8: Administration of neutralizing antibody against ADGRF5 can both prevent and reverse peri-implant fibrosis.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analysed data generated during the study are available for research purposes from the corresponding author within reasonable request. Transcriptomic data from bulk RNA-seq are available from the Gene Expression Omnibus (GEO) under accession numbers GSE227869 and GSE227128. The genomic data used in Fig. 5n and Supplementary Fig. 7 were extracted from publicly available datasets: GSE108892 (ref. 24), GSE138689 (ref. 23) and GSE136970 (ref. 39). The genomic data used in Supplementary Fig. 6 were extracted from the publicly available dataset GSE147287 (ref. 46). Source data for the figures are provided with this paper.

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Acknowledgements

This project was funded by the OREF under awards 994088 and 892405, a Hospital for Special Surgery Surgeon-in-Chief Grant and a Complex Joint Reconstruction Center grant given to V.J.S. X.Y. is supported by grant UL1 TR000457 from the Clinical and Translational Science Center at Weill Cornell Medicine, the Feldstein Medical Foundation and grant W81XWH-21-1-0900 from the Department of Defense. M.B.G. is supported by a Pershing Square Sohn Cancer Research Alliance Award and R01AR075585. The content is solely the responsibility of the authors and does not represent the official views of the sources of research support. We thank E. Monica for the assistance on the article figure editing and formatting. We thank E. Kuyl for the assistance on patient sample procurement. We thank the Flow Cytometry Core, Genomics Resources Core, Optical Microscopy Core and Citigroup Biomedical Imaging Core at Weill Cornell Medicine for their technical support.

Author information

Author notes

  1. These authors contributed equally: Vincentius Jeremy Suhardi, Anastasia Oktarina.

Authors and Affiliations

  1. Department of Orthopedic Surgery, Hospital for Special Surgery, New York, NY, USA

    Vincentius Jeremy Suhardi & Mathias P. G. Bostrom

  2. Research Institute, Hospital for Special Surgery, New York, NY, USA

    Vincentius Jeremy Suhardi, Anastasia Oktarina, Mohammed Hammad, Yingzhen Niu, Qingdian Li, Andrew Thomson, Juan Lopez, Ugur M. Ayturk, Matthew B. Greenblatt, Lionel B. Ivashkiv, Mathias P. G. Bostrom & Xu Yang

  3. Department of Joint Surgery, The Third Hospital of Hebei Medical University, Shijiazhuang, P. R. China

    Yingzhen Niu

  4. Department of Orthopedics, Guangdong Provincial People’s Hospital, Southern Medical University, Guangzhou, P. R. China

    Qingdian Li

  5. Flow Cytometry Core Facility, Weill Cornell Medicine, New York, NY, USA

    Jason McCormick

  6. Department of Orthopedic Surgery, Weill Cornell Medicine, New York, NY, USA

    Ugur M. Ayturk, Mathias P. G. Bostrom & Xu Yang

  7. Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, NY, USA

    Matthew B. Greenblatt

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  1. Vincentius Jeremy Suhardi
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Contributions

V.J.S. and A.O. designed, conducted and analysed the majority of experiments. X.Y., V.J.S., A.O. and M.P.G.B. conceived the project. X.Y. and M.P.G.B. supervised the project. X.Y. performed all mouse surgeries. V.J.S. and A.O. performed μCT scans and analysis and RNA-seq data analysis. V.J.S., J.L. and A.O. maintained and genotyped all mice. J.M. supervised or conducted flow cytometry. V.J.S., A.O., Q.L., M.H., Y.N., Q.L., A.T. and J.L. assisted with surgeries and performed tissue collection, immunofluorescence and histology analysis. M.H. and J.L. helped with human sample processing. M.P.G.B. provided access to human samples and supervised human studies. U.M.A., M.B.G. and L.B.I. helped with the study design and data interpretation. V.J.S., A.O., M.H. and X.Y. prepared the article. All authors read and approved the article.

Corresponding author

Correspondence to Xu Yang.

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The authors declare no competing interests.

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Extended data

Extended Data Fig. 1 LEPR, ACTA2, and ADGRF5 are expressed by peri-implant fibrotic tissue from multiple patients underwent revision surgery for aseptic loosening.

a,c, Expression of LEPR (red), ACTA2(green), ADGRF5 (cyan) by peri-implant fibrotic tissue from two different patients suffering from peri-implant fibrosis around prior total hip arthroplasty. Far left, overlap between DAPI and LEPR. Second from left, overlap between ACTA2 and DAPI. Third from left, overlap between ADGRF5 and DAPI. Third from right, overlap between LEPR and ACTA2. Second from right, overlap between LEPR and ADGRF5. Far right, overlap between LEPR, ACTA2, and ADGRF5. Scale bar, 250 μm. b,d, Expression of LEPR (red), ACTA2 (green), ADGRF5 (cyan) by peri-implant fibrotic tissue from two different patients suffering from peri-implant fibrosis around prior total knee arthroplasty. Far left, overlap between DAPI and LEPR. Second from left, overlap between ACTA2 and DAPI. Third from left, overlap between ADGRF5 and DAPI. Third from right, overlap between LEPR and ACTA2. Second from right, overlap between LEPR and ADGRF5. Far right, overlap between LEPR, ACTA2, and ADGRF5. Scale bar, 250 μm. Images in a–d are representative of 4 biologically independent experiments.

Source data

Extended Data Fig. 2 COL3A1, S100A4, SM22A, ACTA2, FBLN2, and SDC4 are expressed by peri-implant fibrotic tissue in both human and mice.

Expression of COL3A1 (a, green), S100A4 (c, green), SM22A (e, green), ACTA2-RFP (g, green), FBLN2 (i, green), SDC4 (k, green) in murine peri-implant fibrotic tissue and its overlap with LepR– tdTomato cells (red). Scale bar, 50 μm. Expression of COL3A1 (b, green), S100A4 (d, green), SM22A (f, green), ACTA2 (h, green), FBLN2 (j, green), SDC4 (l, green) in human peri-implant fibrotic tissue and its overlap with LEPR+ cells (red). Scale bar, 50 μm. Images in a–l are representative of at least 3 independent biological replicates.

Source data

Extended Data Fig. 3 A subset of LEPR+ cells in the peri-implant fibrotic tissue expresses Cxcl12-GFP.

a, Expression of LepR-tdTomato and Cxcl12-GFP in the peri-implant fibrotic tissue of LepRcre;Rosa26tdTomato; Cxcl12GFP mice underwent fibrous-integrated surgery at postoperative day 14. A subset of LEPR+ cells (red) co-localize with cells expressing Cxcl12 (green). Scale bar, 500 μm. b-e, Enlarged view of the outlined blue box of figure in panel a. Scale bar, 50 μm. Images in a-e are representative of at least 3 independent experiments. f, Expression of LepR-tdTomato and Cxcl12-GFP in the peri-implant osseous tissue of LepRcre;Rosa26tdTomato; Cxcl12GFP mice underwent osseointegrated surgery at postoperative day 14. A subset of LEPR+ cells (red) in the perivascular area co-localizes with cells expressing Cxcl12 (green). Scale bar, 500 μm. g–j, Enlarged view of the outlined blue box of figure in panel f. Scale bar, 50 μm. Images in f-j are representative of at least 3 independent experiments. k–l, Expression of CXCL12-GFP by Lin-LepR-tdTomato+ (k) or osseointegrated surgery (l). m-n, Immunofluorescence quantification of expression of and Cxcl12-GFP in LepRcre;Rosa26tdTomato; Cxcl12GFP mice underwent fibrous-integrated or osseointegrated surgery at postoperative day 14. Data are mean ± s.d. n = 11 for both osseointegrated and fibrous-integrated model. n, Immunofluorescence quantification of co-localization between of LepR-tdTomato and Cxcl12-GFP in LepRcre;Rosa26tdTomato; Cxcl12GFP mice underwent fibrous-integrated or osseointegrated surgery at postoperative day 14. Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. n = 11 for both osseointegrated and fibrous-integrated model. Each dot in m and p corresponds to biologically independent replicates.

Source data

Extended Data Fig. 4 Small percentage of Lin– 6C3– CD90–CD200+ CD105– (mSSC) and Lin– 6C3–CD90– CD200var CD105+ (BCSP) are Lin– LepR– tdTomato+.

a, Schematic of flow cytometry comparison experiment between LepRcre;Rosa26tdtomato mice underwent fibrous-integrated or osseointegrated implantation surgery at postoperative day 14. b-d, Schematic representation of the strategy used for FACS analysis of Lin– 6C3– CD90– CD200+ CD105– (mSSC) and Lin-6C3-CD90-CD200varCD105+ (BCSP)of mice underwent fibrous integrated surgery (c) or osseointegrated surgery (d). e–f, There is no difference in abundance of mSSC and BCSP in the peri-implant area between fibrous- and osseointegrated mice. Data are mean ± s.d. Unpaired, two-tailed Student’s t-test. n = 8 for fibrous-integrated model and n = 9 for osseointegrated model. g, TdTomato+ subset of Lin– 6C3– CD90– CD200+ CD105– are more abundant in fibrous-integrated peri-implant area than in osseointegrated peri-implant area. Data are mean ± s.d. Unpaired, two-tailed Student’s t-test. n = 8 for fibrous-integrated model and n = 9 for osseointegrated mode. h, There is no statistically significant difference in the abundance of tdTomato+ subset of Lin-6C3– CD90– CD200var CD105+ in fibrous-integrated peri-implant and in osseointegrated peri-implant area. Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. n = 8 for fibrous-integrated model and n = 9 for osseointegrated model. i, LepR– tdTomato+ cells in fibrous-integrated interface were immunostained for CD200 demonstrating presence of LepR-TdTomato+ CD200+ at the bone-fibrous tissue interface. Scale bar, 500 μm. Middle column and right column enlarged view of the outlined yellow box. Scale bar, 50 μm. j, LepR– tdTomato+ cells in osseointegrated interface were immunostained for CD200 demonstrating presence of LepR– TdTomato+ CD200+ at the perivascular area. Scale bar, 500 μm. Middle column and right column, enlarged view of the outlined yellow box. Scale bar, 50 μm. Each dot in e–h corresponds to biologically independent replicates. Images in b–d, i–j are representative of at least 3 independent experiments.

Source data

Extended Data Fig. 5 A subset of LEPR+ cells in the peri-implant.

a, Expression of LepR-zsGreen and Acta2-RFP in the peri-implant fibrotic tissue of LepRcre;Rosa26Zsgreen; Acta2mRFP mice underwent fibrous-integrated surgery at postoperative day 14. A subset of LEPR+ cells (green) co-localize with cells expressing Acta2 (red). Scale bar, 500 μm. b-e, Enlarged view of the outlined yellow box of the figure in panel a. Scale bar, 50 μm. Images in a-e are representative of at least 3 independent experiments. f, Expression of LepR-zsGreen and Acta2-RFP in the peri-implant osseous tissue of LepRcre;Rosa26Zsgreen; Acta2mRFP mice underwent osseointegrated surgery at postoperative day 14. A subset of LEPR+ cells (green) in the perivascular area co-localize with cells expressing Acta2 (red). Scale bar, 500 μm. Images in a are representative of at least 3 independent experiments. g–j, Enlarged view of the outlined yellow box of figure in panel f. Scale bar, 50 μm. Images in f–j are representative of at least 3 independent experiments. k–l, Representation of the strategy used for FACS analysis of expression of LepR-zsGreen and Acta2-RFP in Lin-6C3-CD90-CD200+ CD105– fibrous integrated surgery (k) or osseointegrated surgery (l). LepRcre;Rosa26Zsgreen; Acta2mRFP (red), LepRcre;Rosa26Zsgreen (blue). m-n, Immunofluorescence quantification of expression of LepR-zsGreen and Acta2-RFP in LepRcre;Rosa26Zsgreen; Acta2mRFP mice underwent fibrous-integrated or osseointegrated surgery at postoperative day 14. Data are mean ± s.d. n = 5 for both osseointegrated and fibrous-integrated model. Unpaired, two-tailed Student’s t-test. o, Immunofluorescence quantification of co-localization between LepR-zsGreen and Acta2-RFP in LepRcre;Rosa26Zsgreen; Acta2mRFP mice underwent fibrous-integrated or osseointegrated surgery at postoperative day 14. Data are mean ± s.d. n = 5 for both osseointegrated and fibrous-integrated model; Unpaired, tailed Student’s t-test. p, Flow cytometry quantification of relative abundance of Lin-LepR-Zsgreen+ Acta2-RFP+ 6C3–CD90– CD200+ CD105 in LepRcre;Rosa26Zsgreen; Acta2mRFP mice underwent fibrous-integrated or osseointegrated surgery at postoperative day 14. Data are mean ± s.d. n = 5 for both osseointegrated and fibrous-integrated model; Unpaired, two-tailed Student’s t-test. Each dot in m-p corresponds to biologically independent replicates. Images in k-l are representative of at least 3 independent experiments.

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Extended Data Fig. 6 Peri-implant fibrotic tissue originate from osteogenic LEPR+ cells.

a–c, Immunofluorescent imaging of proximal tibia of LepRcre;Rosa26tdtomato, BGLAP-GFP that underwent fibrous-integration surgery at post-operative day 3 (a), 7 (b), and 14 (c). Scale bar, 50 μm. d–f, Immunofluorescent imaging of ALPL-stained proximal tibia of LepRcre;Rosa26tdtomato, BGLAPGFP that underwent fibrous-integration surgery at post-operative day 3 (d), 7 (e), and 14 (f). Scale bar, 50 μm. g, Violin plots showing the expression density levels of Alpl and Bglap showed that they are selectively expressed by osteogenic (OLC1, OLC2) and chondrogenic (CLC) cells. h, Immunofluorescence quantification of co-localization between LepR-tdTomato and BGLAP-GFP in LepRcre;Rosa26tdtomato, BGLAPGFP mice underwent fibrous-integrated surgery at postoperative day 3, 7, and 14. Data are mean ± s.d. n = 6 for both osseointegrated and fibrous-integrated model; Unpaired, tailed Student’s t-test. i, Immunofluorescence quantification of co-localization between LepR-tdTomato and ALPL antibody staining in LepRcre;Rosa26tdtomato, BGLAPGFP mice underwent fibrous-integrated surgery at postoperative day 3, 7, and 14. Data are mean ± s.d. n = 6 for both osseointegrated and fibrous-integrated model; Unpaired, tailed Student’s t-test. j, Immunofluorescence quantification of tdTomato+ cells from CXCL12creERT2;Rosa26tdtomato, AdipoqcreERT2;Rosa26tdtomato and LepRcre;Rosa26tdtomato that underwent fibrous-integration surgery. N = 7 per group. Data are mean ± s.d.; two-tailed unpaired t-test. k-m, Immunofluorescent imaging of CXCL12creERT2;Rosa26tdtomato (k), AdipoqcreERT2;Rosa26tdtomato (l), and LepRcre;Rosa26tdtomato (m) that underwent fibrous-integration surgery at post-operative day 14. Each dot in h-j corresponds to biologically independent replicates. Images in a–f, k–m are representative of at least 3 independent experiments.

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Extended Data Fig. 7 Peri-implant fibrosis is persistent up to at least 16 weeks in murine model of peri-implant fibrosis.

a, Micro-computed tomography (μCT) of proximal tibia of LepRcre;Rosa26tdtomato underwent fibrous-integrated surgery at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. b, Hematoxylin and eosin staining of proximal tibia of LepRcre;Rosa26tdtomato underwent fibrous-integrated surgery at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. c, Immunofluorescent imaging proximal tibia of LepRcre;Rosa26tdtomato underwent fibrous-integrated surgery at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. Bottom row, enlarged view of the outlined yellow box at each time-point. Scale bar, 50 μm. d, Bone volume/total volume (BV/TV)) of mice at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6. e, Histology quantification of peri-implant fibrosis (% Fibrosis) and peri-implant bone (% bone) of mice at at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6. f, Immunofluorescent quantification of peri-implant fibrosis (% Fibrosis) and peri-implant bone (% bone) of mice at at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6. Each dot in d-f corresponds to biologically independent replicates. Images in a–c are representative of at least 3 independent experiments.

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Extended Data Fig. 8 Analysis of peri-implant bone of mice underwent osseointegrated surgery at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks.

a, Micro-computed tomography (μCT) of proximal tibia of LepRcre;Rosa26tdtomato underwent osseointegrated surgery at post-operative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. b, Haematoxylin and eosin staining of proximal tibia of LepRcre;Rosa26tdtomato underwent osseointegrated surgery at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. c, Immunofluorescent imaging proximal tibia of LepRcre;Rosa26tdtomato underwent osseointegrated surgery at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Scale bar, 500 μm. Bottom row, enlarged view of the outlined yellow box at each time-point. Scale bar, 50 μm. d, Bone volume/total volume (BV/TV) of mice at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6. e, Histology quantification of peri-implant fibrosis (% Fibrosis) and peri-implant bone (% bone) of mice at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6; 3 independent experiments. f, Immunofluorescent quantification of peri-implant fibrosis (% Fibrosis) and peri-implant bone (% bone) of mice at postoperative day 3, day 7, 2 weeks, 4 weeks, 8 weeks, and 16 weeks. Data are mean ± s.d. n = 6. Each dot in d-f corresponds to biologically independent replicates. Images in a-c are representative of at least 3 independent experiments.

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Extended Data Fig. 9 Human mSSC (Lin– PDPN+ CD146– CD164+ CD73+) are more abundant in bone than in fibrous membranes, but Lin– LEPR+ are more abundant in fibrous membranes than in bone.

a, Schematic representation of the strategy used for FACS analysis to obtain Lin- population. b–c, Schematic representation of the strategy used for FACS analysis to obtain LEPR expressing a subset of Lin– PDPN+ CD146– CD164+ CD73+ from bone (b) or from fibrous tissue (c). d, Flow cytometry quantification of human fibrous tissue and bone demonstrates human mSSC (Lin– PDPN+ CD146– CD164+ CD73+) are more abundant in bone than in fibrous membrane. Data are mean ± s.d. n = 10 patients for fibrous membrane and n = 10 for bone patients. Unpaired, two-tailed Student’s t-test. e, Flow cytometry quantification of human fibrous tissue and bone demonstrates the non-significant difference in osteolineage progenitor (Lin–PDPN+ CD146+) between fibrous membrane and bone. Data are mean ± s.d. n = 10 for fibrous membrane and n = 10 for bone. Unpaired, two-tailed Student’s t-test. f, Flow cytometry quantification of human fibrous tissue and bone demonstrates significantly higher amount of Lin– LEPR+ PDPN+ CD146– in the fibrous membrane than in bone. Data are mean ± s.d. n = 10 patients for fibrous membrane and n = 10 patients for bone. Unpaired, two-tailed Student’s t-test. Each dot in d-f corresponds to biologically independent replicates. Images in a-c¬ are representative of at least 3 independent experiments.

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Extended Data Fig. 10 Administration of neutralizing antibody against reduces LepR– tdTomato+ cells but does not affect Lin– 6C3– CD90– CD200+ CD105– (mSSC) subpopulation.

a, Immunofluorescent imaging of CD200-antibody stained. proximal tibia of LepRcre;Rosa26tdtomato receiving either anti-ADGRF5 or isotype control as prophylaxis. Scale bar, 500 μm. Right column, enlarged view of the outlined yellow box. Scale bar, 50 μm. b, Immunofluorescent imaging of CD200-antibody-stained proximal tibia of LepRcre;Rosa26tdtomato receiving either anti-ADGRF5 or isotype control as treatment starting from postoperative day 14. Scale bar, 500 μm. Right column, enlarged view of the outlined yellow box. Scale bar, 50 μm. c,f FACS analysis of TdTomato+ subset of Lin-6C3– CD90–CD200+ CD105– (TdTomato+ mSSC) in mice group receiving anti-ADGRF5 as compared to isotype control, either as prophylaxis (c) or as treatment (f). Anti-ADGRF5 (n = 5), isotype control (n = 5). Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. d,g, FACS analysis of Lin– 6C3– CD90– CD200+ CD105– (mSSC) between mice receiving daily anti-ADGRF5 or isotype control either as prophylaxis (d) or as treatment (f). Anti-ADGRF5 (n = 5), isotype control (n = 5). ns= not significant. Unpaired, two-tailed Student’s t-test. treatment (f). Anti-ADGRF5 (n = 5), isotype control (n = 5). Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. e, Host bone-implant failure load of LepRcre;Rosa26tdtomato receiving either daily anti-ADGRF5 or isotype control as prophylaxis at postoperative day 28. n = 5 for mice receiving anti-ADGRF5 and n = 5 for mice receiving isotype control. Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. h, Host bone-implant failure load of LepRcre;Rosa26tdtomato receiving either daily anti-ADGRF5 or isotype control as treatment starting from postoperative day 14 and evaluated at postoperative day 28. n = 5 for mice receiving anti-ADGRF5 and n = 5 for mice receiving isotype control. Unpaired, two-tailed Student’s t-test. Data are mean ± s.d. Images in a-b are representative of at least 3 independent biological replicates. Each dot in c–e and f–h corresponds to biologically independent replicates.

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Suhardi, V.J., Oktarina, A., Hammad, M. et al. Prevention and treatment of peri-implant fibrosis by functionally inhibiting skeletal cells expressing the leptin receptor. Nat. Biomed. Eng 8, 1285–1307 (2024). https://doi.org/10.1038/s41551-024-01238-y

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