Optimizing Operational Parameters to Preserve Cellular Integrity During Diagnostic Vitrectomy: An Ex-Vivo Pilot Study With Blinded Outcome Assessment
试验速览
- 阶段
- 不适用
- 状态
- 尚未招募
- 入组人数
- 100
- 主要终点
- Difference in absolute lymphocyte count between post-processing and baseline measurements (Δ lymphocyte count).
研究概览
简要总结
Single-center ex-vivo study using blood samples from healthy donors processed through a 25-gauge vitrectomy system. The study compares four combinations of operational parameters (vacuum pressure and cut rate) to evaluate their impact on cellular recovery and morphological preservation. Pre- and post-procedure complete blood counts and cytology smears will be analyzed to identify the settings that minimize cell loss and mechanical damage, with the ultimate goal of optimizing diagnostic yield in vitreoretinal lymphoma (VRL) vitrectomy.
详细描述
Primary vitreoretinal lymphoma (VRL) is a rare but aggressive intraocular malignancy, typically classified as a subtype of primary central nervous system lymphoma. It predominantly involves the retina and vitreous and is frequently associated with central nervous system disease. Clinical presentation often mimics chronic uveitis, leading to diagnostic delays and suboptimal management.
Definitive diagnosis of VRL requires cytopathologic confirmation through diagnostic vitrectomy, which provides vitreous samples for cytology, immunohistochemistry, flow cytometry, and molecular analyses (e.g., MYD88 mutation testing, IL-10/IL-6 ratio). However, despite the central role of vitreous biopsy, its diagnostic sensitivity remains limited to approximately 70% (Iuliano L et al. Int Ophthalmol 43, 2841-2849 (2023)), primarily due to the mechanical fragility and scarcity of malignant lymphoid cells.
During vitrectomy, the vitreous is fragmented and aspirated through a high-speed cutting probe (vitrector), which applies both mechanical cutting and vacuum forces. These operational parameters, while optimized for surgical efficiency and visualization, may inadvertently damage cells by rupturing membranes or disrupting nuclei, thereby reducing the yield and integrity of cellular material available for analysis.
Given the critical importance of maximizing diagnostic sensitivity in a disease with high morbidity and mortality, refining vitrectomy parameters to minimize cellular damage could represent a major advancement in the diagnostic workflow of VRL.
The present study aims to systematically evaluate the effect of different combinations of vacuum pressure and cut rate on cellular recovery and morphological preservation in a controlled ex-vivo vitreous model. Approximately 100 healthy donors will be enrolled and stratified into four experimental groups, each undergoing simulated diagnostic vitrectomy procedures under distinct parameter settings:
研究设计
- 研究类型
- Observational
- 观察模型
- Other
- 时间视角
- Cross Sectional
入排标准
- 年龄范围
- 18 Years 至 65 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Provision of written informed consent authorizing the anonymized research use of residual blood samples.
- •Age 18-65 years at the time of donation.
- •Eligibility for routine whole blood donation, according to the institutional and national standards of the Blood Transfusion Center.
- •Good general health, as per standard donor screening procedures.
排除标准
- •No additional exclusion criteria are defined beyond standard blood donation regulations. Exclusion from the study will therefore coincide with ineligibility for blood donation according to institutional protocols (e.g., due to infection, hematologic disease, recent surgery, or medication use that affects blood composition).
研究组 & 干预措施
HV/LC
High vacuum (650 mmHg) / Low cut rate (800 cpm)
干预措施: Simulated vitrectomy (Other)
LV/HC
Low vacuum (300 mmHg) / High cut rate (10,000 cpm)
干预措施: Simulated vitrectomy (Other)
LV/LC
Low vacuum (300 mmHg) / Low cut rate (800 cpm)
干预措施: Simulated vitrectomy (Other)
HV/HC
High vacuum (650 mmHg) / High cut rate (10,000 cpm)
干预措施: Simulated vitrectomy (Other)
结局指标
主要结局
Difference in absolute lymphocyte count between post-processing and baseline measurements (Δ lymphocyte count).
时间窗: Same-day assessment, immediately after processing
To quantify the impact of different vitrectomy parameter combinations (vacuum pressure and cut rate) on lymphocyte recovery after ex-vivo processing of whole blood using a vitreous trap model.
次要结局
- Δ values for the listed CBC parameters(Same-day assessment, immediately after processing)
- Semiquantitative cytology integrity score (smear evaluation)(Same-day assessment, immediately after processing)