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This study aims to explore the dynamic changes in different cell types and their molecular regulatory mechanisms during the healing process of peri-implant soft tissues, using single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST).
The research will focus specifically on the healing of soft tissues around dental implants in human subjects with the use of a healing abutment. Participants in the healing abutment group will have a micro healing abutment placed immediately after implant placement.
Tissue samples will be collected at various time points (Day 4, Day 14, and Day 70) to construct a detailed single-cell map of the healing process.
Spatial transcriptomics will be integrated to preserve tissue architecture, enabling the identification of the spatial distribution of different cell types and their interactions within the peri-implant microenvironment.
This combined approach will allow for a comprehensive characterization of cellular interactions, spatial gene expression patterns, and regulatory networks specific to healing abutments.
The goal is to identify key regulatory factors that could improve peri-implant soft tissue healing and help prevent peri-implantitis.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Healing Abutment Gingival Healing Post-Implantation | Experimental |
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| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Healing Abutment Gingival Healing Post-Implantation | Procedure | The healing process of peri-implant soft tissues will be studied using a micro healing abutment (2 mm), which will be placed at least three months after implant Stage I surgery. Gingival tissue samples will then be collected at Day 4, Week 2, Week 4, and Week 8-10. At each time point, a circular incision around the micro abutment will be made using a circular punch, and the gingival tissue together with the abutment will be retrieved under local anesthesia. After sampling, the micro abutment will be replaced with a formal healing abutment to maintain healing. Samples will undergo single-cell RNA sequencing (scRNA-seq) to characterize cell types, gene expression changes, and regulatory pathways involved in soft tissue repair. Additional tissues will be used for spatial transcriptomics (ST) and histological staining. |
| Measure | Description | Time Frame |
|---|---|---|
| Primary Outcome 1: Differentially Expressed Genes (DEGs) in Peri-implant Soft Tissues | Number of genes significantly differentially expressed between experimental groups, determined by single-cell RNA sequencing (scRNA-seq) analysis (FDR < 0.05). Unit of Measure: Number of genes. | Day 4, Week 2, Week 4 and Week 8-10 post-implantation. |
| Cell Type Composition in Peri-implant Soft Tissues | Proportion (%) of each annotated cell type identified by scRNA-seq based on canonical marker expression. Unit of Measure: Percentage of total cells (%) | Day 4, Week 2, Week 4, and Week 8-10 post-implantation. |
| Enriched Biological Pathways | Number of significantly enriched Gene Ontology (GO) terms and KEGG pathways identified from DEGs (FDR < 0.05). Unit of Measure: Number of enriched terms/pathways | Day 4, Week 2, Week 4, and Week 8-10 post-implantation |
| Ligand-Receptor Interaction Strength | Mean interaction score of ligand-receptor pairs identified among annotated cell types using CellPhoneDB (or equivalent computational tool). Unit of Measure: Interaction score (arbitrary units) | Day 4, Week 2, Week 4, and Week 8-10 post-implantation |
| Spatial Gene Expression Patterns | Localization and expression intensity of selected DEGs across tissue regions determined by the 10x Genomics Visium platform. | Day 4, Week 2, Week 4 and Week 8-10 post-implantation. |
| Measure | Description | Time Frame |
|---|---|---|
| Histological Tissue Morphology | Assessment of epithelial continuity, connective tissue organization, and inflammatory cell infiltration in peri-implant soft tissues by hematoxylin and eosin (H&E) staining. Each parameter will be semi-quantitatively scored on a standardized histological scale (0-3), where 0 = none, 1 = mild, 2 = moderate, and 3 = severe. | Day 4, Week 2, Week 4 and Week 8-10 post-implantation. |
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Inclusion Criteria:
Exclusion Criteria:
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| Name | Role | Phone | Extension | |
|---|---|---|---|---|
| Hui Cheng | Contact | +86 18960883888 | ch_fujian@163.com |
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| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| School and Hospital of Stomatology, Fujian Medical University | Fuzhou | Fujian | 350004 | China |
Taking into account factors such as the needs of subsequent research, the final decision has not yet been made.
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| Immune and Stromal Cell Quantification | Quantification of immune cells (e.g., CD68⁺ macrophages, CD3⁺ T lymphocytes) and stromal cells (e.g., fibroblast-related markers) by immunohistochemistry (IHC). Mean positive cell count per high-power field (HPF) will be calculated from representative sections. | Day 4, Week 2, Week 4 and Week 8-10 post-implantation. |
| Junctional Epithelium Length | Measurement of junctional epithelium length along the implant surface using histological sections and digital image analysis software. Unit of Measure: Length (µm) | Day 4, Week 2, Week 4, and Week 8-10 post-implantation. |
| Basement Membrane Protein Expression | Immunohistochemical detection of Laminin-332 and related basement membrane proteins indicating epithelial attachment integrity. The percentage of positive staining area will be quantified using image analysis software. Unit of Measure: Percentage of positive area (%) | Day 4, Week 2, Week 4, and Week 8-10 post-implantation. |