The Role of Type H Vessel Formation in Induced Membrane of Patients With Critical Size Bone Defect

March 30, 2023 updated by: Chang Gung Memorial Hospital

The Role of Endothelial Cell-mediated Type H Vessel Formation in Induced Membrane-enhanced Bone Union of Patients With Critical Size Bone Defect

The goal of this clinical trial is to test the role of type H vessel and investigate the mechanism of its regulation in induced membrane created by induced membrane technique in patients with open fracture, aged between 20 and 80.

The main questions to answer are:

  • Whether type H vessel will be critical for induced membrane-mediated bone healing?
  • Whether type H vessel will be a biomarker for diagnosing bone healing in patients with open fracture?
  • To find out a circulating indicator for type H vessel in patients with open fracture undergoing induced membrane technique.

Participants will be enrolled to collect tissues of induced membrane and blood samples to detect type H vessel and measure the biomarkers of type H vessel, investigating their correlation with the capacity of bone healing. To avoid selection bias of treatment, the grouping will be achieved by a randomized protocol qualified clinical trial center of Chang Gung Memorial Hospital to divide into with induced membrane and without induced membrane. Researchers will compare these two groups to test our hypothesis that type H vessel in important in induced membrane and their corresponding biomarkers.

Study Overview

Status

Not yet recruiting

Conditions

Intervention / Treatment

Detailed Description

The masquelet technique, also called an induced membrane technique (IMT), was first introduced to treat critical size bone defect up to 25 cm in 1986 by Pr. Alain Charles Masquelet et al. IMT is a two-stage surgical operation that recently provides a more popular option for the treatment of critical size bone defect as well as fracture nonunion. Basically, the first stage of IMT comprises the implantation of a polymethylmethacrylate (PMMA) cement spacer loaded with antibiotics into defect site. During 6 to 8 weeks, this spacer induces the formation and maturation of a thin layer called induced membrane surrounding it via a foreign body immune response. In stage II, the spacer is carefully removed while keeping induced membrane at the fracture site, autologous bone graft is then performed to provoke bone healing. IMT endows several beneficial effects including the relatively simple management versus vascularized fibular grafts and no significant difference in healing time among various defect sizes. Of note, multiple surgeries and long recovery time are the difficulties still needed to be conquered in patients treated with IMT. In view of the fact that induced membrane has been considered as an indispensable component for successful union, a more detailed and comprehensive analysis of it will improve the process of IMT to ameliorate those difficulties.

The clinical study design will follow the description in Chang Gung Medical Foundation Institutional Review Board, which will be begun in the first year until reaching 15 patients for each group. Briefly, patients of open fracture will be randomly divided into two groups to reduce selection bios, comprising spacer (with induced membrane) and without spacer groups (without induced membrane). In a without spacer group (No operation of stage I), soft tissues near fracture sites with a size of 1 cm2 will be collected to be used as an appropriate control of induced membrane (1 cm2) from the spacer group (Stage I + II) at the time point of stage II of IMT. Tissues will be dissected into three parts for proteomics, metabolomics, and paraffinized section, stored at minus 80 degrees prior to experiments. The section is imperative in this project to characterize type H vessel formation and the capacity of bone healing. For this purpose, immunohistochemistry (IHC) or Immunofluorescence (IF) will be performed to measure identified proteins from proteomics results in both of soft tissues and induced membrane. The characterized markers comprising cluster of differentiation 31 (CD31) and Endomucin (Emcn) will be used to detect type H vessels. Colorimetric methods including Masson's trichrome, Alcian blue, Alizarin red S, H&E and Safranin O-Fast green can also be performed to unveil histological insights of induced membrane. In the multi-omics, tissues will be used to unveil novel targets, which will then be compared to the levels of vascular endothelial growth factor (VEGF) and osteocalcin. Furthermore, western blotting and real-time reverse transcriptase-Polymerase Chain Reaction (RT-PCR) will be performed to validate downstream regulation of the identified metabolites.

The venous blood (3ml in vacutainers with heparin) will be collected from both groups at the Department of Orthopedic Surgery, Chang Gung Memorial Hospital, Taoyuan, Taiwan. The protocol was based on a report identifying VEGF as a potential biomarker of the union in IMT. After drawing, blood will be centrifuged (1000RPM, 10min at room temperature) for 30 minutes and stored at minus 80 degrees until analysis. Blood samples from the time point of collecting induced membrane are prepared for multi-omics analysis to identify systemic markers for type H vessel or induced membrane, and VEGF and osteocalcin will be used as serum indicators for bone mass. Blood samples from all time points will be used to identify a novel biomarker for type H vessels and bone healing. As for metabolite targets, targeted methods of metabolomics will be employed to verify them in blood. The time points for collecting blood samples of stage I will be the day of surgery, the next day of surgery, and 1, 2, and 4 weeks after surgery. As for stage II, the time points will be the day of surgery, the next day of surgery, and 1, 2, 12, and 14 weeks after surgery.

Study Type

Interventional

Enrollment (Anticipated)

30

Phase

  • Not Applicable

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

  • Name: HSU YUNG-HENG, MD, PhD
  • Phone Number: 2423 (03)3281200
  • Email: hsuyh@cgmh.org.tw

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

20 years to 80 years (Adult, Older Adult)

Accepts Healthy Volunteers

No

Genders Eligible for Study

All

Description

Inclusion Criteria:

  • Accept the informed consent form
  • Age: 20 years to 80 years
  • Type II and III open fracture with 2-6.5 cm defect size

Exclusion Criteria:

  • Not accept the informed consent form
  • Age smaller than 20 years and over 80 years
  • Quit during the trial
  • patients with Notifiable infectious diseases such as AIDS and hepatitis as well as cancer

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Primary Purpose: Basic Science
  • Allocation: Randomized
  • Interventional Model: Parallel Assignment
  • Masking: Double

Arms and Interventions

Participant Group / Arm
Intervention / Treatment
Experimental: with induced membrane
operation of stage I and II
Procedure: induced membrane technique
Masquelet technique, also called an induced membrane technique (IMT), was first introduced to treat critical size bone defect up to 25 cm in 1986 by Pr. Alain Charles Masquelet et al. IMT is a two-stage surgical operation that recently provides a more popular option for the treatment of critical size bone defect as well as fracture nonunion. Basically, first stage of IMT comprises the implantation of a polymethylmethacrylate (PMMA) cement spacer loaded with antibiotics into defect site. During 6 to 8 weeks, this spacer induces the formation and maturation of a thin layer called induced membrane surrounding it via a foreign body immune response. In stage II, the spacer is carefully removed while keeping induced membrane at the fracture site, autologous bone graft is then performed to provoke bone healing.
Other Names:
  • Masquelet technique
Experimental: without induced membrane
only stage II
Procedure: Bone graft (Stage II of IMT)
Bone graft is only performed in this group to provoke bone healing, as same as demonstrated in the stage II of IMT.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
The participants with open fracture will be assessed by the level of type H vessel
Time Frame: Induced membrane will be collected at 8th weeks of IMT performing procedures of stage II.
In each group, induced membrane and soft tissue collected at 8th weeks will be used to detect CD31 and Emcn by performing immunohistochemistry (IHC) or immunofluorescence (IF), two markers for identifying type H vessel that correlated to the capacity of bone healing. The staining will be quantified as units of intensity in values defined by software or area in pixels to assess the level of type H vessel in each participant.detecting the expression level in induced membrane and soft tissue at 8 weeks, and markers of bone healing as well as metabolites in blood of each point.
Induced membrane will be collected at 8th weeks of IMT performing procedures of stage II.
The participants with open fracture will be assessed by X-ray
Time Frame: Bone healing tracking will be up to 24 weeks after stage II operation.
The defect sites in these patients will be assessed the capacity of bone healing in a time-dependent manner, by quantifying the sizes of the defects from these images and representing them as the length in centimeter (cm) or area in cm2. These results will be aggregated with the level of type H vessel to report the correlation between bone healing and type H vessel.
Bone healing tracking will be up to 24 weeks after stage II operation.
The participants with open fracture will be assessed the metabolic alteration
Time Frame: Induced membrane will be collected at 8th weeks of IMT performing procedures of stage II. Blood will be a time-dependent manner through out this study up to 32 weeks.
Finding out a potential marker to predict the correlation mentioned above, bloods collected from patients in a time-dependent manner and tissue at 8th week will be assessed the metabolic alteration by performing analysis of Liquid chromatography-mass spectrometry (LC-MS/MS). The identified metabolites will be represented as a unit of concentration in micro molar. The levels of metabolites will also be aggregated with results of type H vessel and bone healing to report whether these identified metabolites will be a type H marker defining or predicting the capacity of bone healing.
Induced membrane will be collected at 8th weeks of IMT performing procedures of stage II. Blood will be a time-dependent manner through out this study up to 32 weeks.

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Chang Gung Memorial Hospital

Investigators

  • Principal Investigator: HSU YUNG-HENG, MD, PhD, Chang Gung Memorial Hospital

Publications and helpful links

The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.

General Publications

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Anticipated)

April 1, 2023

Primary Completion (Anticipated)

April 1, 2023

Study Completion (Anticipated)

January 2, 2026

Study Registration Dates

First Submitted

February 15, 2023

First Submitted That Met QC Criteria

March 30, 2023

First Posted (Actual)

March 31, 2023

Study Record Updates

Last Update Posted (Actual)

March 31, 2023

Last Update Submitted That Met QC Criteria

March 30, 2023

Last Verified

January 1, 2023

More Information

Terms related to this study

Keywords

Other Study ID Numbers

  • 202201532B0A3

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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