Immunophenotyping of Fresh Stromal Vascular Fraction From Adipose Derived Stem Cells (ADSC) Enriched Fat Grafts

• Sponsor: University of Sao Paulo (Other)
• Overall Status: Completed
• CT.gov ID: NCT01771913
• Collaborator: (none)
• Enrollment: 20
• Location: 1
• Arms: 2
• Duration (Months): 38
• Patients Per Site Per Month: 0.5

Study Details

Study Description

Brief Summary

The purpose of this study is to investigate if there is a relationship between the take of fat grafts with and without ADSCs and the presence of specific surface markers on the cells of the stromal vascular fraction.

Condition or Disease	Intervention/Treatment	Phase
Breast Reconstruction; Contour Irregularities; Volume Insufficiency	Genetic: centrifuged fat graft; Genetic: ADSCs enriched fat graft	Phase 2

Detailed Description

Two groups of patients will be studied. The control will receive centrifuged fat grafts while the other group will receive centrifuged enriched fat grafts with ADSCs. The performance of both grafts will be assessed through volume measurement employing MRI before and after 6 months post-op.

Study Design

• Study Type: Interventional
• Actual Enrollment: 20 participants
• Allocation: Randomized
• Intervention Model: Parallel Assignment
• Masking: Double (Participant, Outcomes Assessor)
• Primary Purpose: Treatment
• Official Title: Immunophenotyping of Fresh Stromal Vascular Fraction From Adipose Derived Stem Cells (ADSC) Enriched Fat Grafts for Refinements of Reconstructed Breasts
• Study Start Date: Mar 1, 2012
• Actual Primary Completion Date: Sep 1, 2014
• Actual Study Completion Date: May 1, 2015

Arms and Interventions

Arm	Intervention/Treatment
Sham Comparator: centrifuged fat graft female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements.	Genetic: centrifuged fat graft; fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.
Active Comparator: ADSCs enriched centrifuged fat graft female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement	Genetic: ADSCs enriched fat graft; fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.

Outcome Measures

Primary Outcome Measures

1. Volume Maintenance [up to 1 year]

   Volumetry of the reconstructed breasts will be accomplished through MRI and OsiriX software. Osirix software allows breast volume calculation through the determination of regions of interest (ROIs) on an MRI sequence. Once the pre (V1) and postoperative (V2) volumes were determined the following formula was applied: (V2 - V1) X 100/graft volume. The result, expressed in percentage, expresses graft volume persistence.

Secondary Outcome Measures

1. Immunophenotyping [baseline]

   Immunophenotyping of the fresh stromal vascular fraction of both groups. Immunophenotyping or flow cytometry measures how many cells, in a sample, express a specific surface marker. A surface marker or a group of markers may characterize a specific cell type. The software that accompanies the flow cytometer determines the number of cells (in percentage) that express the tested surface marker.

Other Outcome Measures

1. Number of Participants Experiencing Fat Necrosis in the Postoperative Period [up to 3 years]

   Fat necrosis may occur whenever a fat graft is performed and it has clinical relevance. It can emerge as oil cysts or small nodules a little bit painful. In mammograms of normal breasts, fat necrosis present as cysts or micro calcifications that present a benign appearance. In breast reconstruction patients, fat necrosis, despite its benign characteristics, can suggest cancer recurrence.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 20 Years to 70 Years
• Sexes Eligible for Study: Female
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• contour irregularities and volume insufficiency in reconstructed breasts no matter the method of reconstruction employed

• local flaps with conditions to receive fat grafts

• good health condition

Exclusion Criteria:

• breast cancer patients under chemotherapy

• smokers

• bad health condition

• patients too thin

• patients that require a new reconstructive surgery (secondary reconstruction)

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Hospital Municipal Carmino Caricchio	Sao Paulo		Brazil	03063-000

Sponsors and Collaborators

• University of Sao Paulo

Investigators

• Study Chair: Nivaldo Alonso, PhD, University of Sao Paulo

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

• Responsible Party: Luiz Alexandre Lorico Tissiani, MD, Chief of Breast Reconstructive Unit at Hospital Municipal Carmino Caricchio, University of Sao Paulo
• ClinicalTrials.gov Identifier: NCT01771913
• Other Study ID Numbers: boosteredADSCs
• First Posted: Jan 18, 2013
• Last Update Posted: Aug 24, 2015
• Last Verified: Jul 1, 2015

• Keywords provided by Luiz Alexandre Lorico Tissiani, MD, Chief of Breast Reconstructive Unit at Hospital Municipal Carmino Caricchio, University of Sao Paulo: adipocytes; cell transplantations; mesenchymal stem cell; immunophenotyping; adipose tissue; breast reconstruction; mammaplasty; magnetic resonance imaging; fat necrosis

Study Results

Participant Flow

Recruitment Details	patients who have contour irregularities after any method of breast reconstruction. recruited from jan/2012 to august/2014.

Arm/Group Title	Centrifuged Fat Graft	ADSCs Enriched Centrifuged Fat Graft
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.	female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.
Period Title: Overall Study
STARTED	9	11
COMPLETED	8	11
NOT COMPLETED	1	0

Baseline Characteristics

Arm/Group Title	Centrifuged Fat Graft	ADSCs Enriched Centrifuged Fat Graft	Total
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.	female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.	Total of all reporting groups
Overall Participants	8	11	19
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	7 87.5%	11 100%	18 94.7%
>=65 years	1 12.5%	0 0%	1 5.3%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	49.8 (11.5)	49.6 (5.7)	49.7 (8.3)
Sex: Female, Male (Count of Participants)
Female	8 100%	11 100%	19 100%
Male	0 0%	0 0%	0 0%
Region of Enrollment (participants) [Number]
Brazil	8 100%	11 100%	19 100%
breast volume (mL) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [mL]	562.8 (187.7)	650.1 (206.8)	613.3 (198.5)

Outcome Measures

1. Primary Outcome

Title	Volume Maintenance
Description	Volumetry of the reconstructed breasts will be accomplished through MRI and OsiriX software. Osirix software allows breast volume calculation through the determination of regions of interest (ROIs) on an MRI sequence. Once the pre (V1) and postoperative (V2) volumes were determined the following formula was applied: (V2 - V1) X 100/graft volume. The result, expressed in percentage, expresses graft volume persistence.
Time Frame	up to 1 year

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Centrifuged Fat Graft	ADSCs Enriched Centrifuged Fat Graft
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.	female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.
Measure Participants	8	11
Mean (Standard Deviation) [percentage of graft volume persistency]	51.6 (18.4)	78.9 (74.9)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Centrifuged Fat Graft, ADSCs Enriched Centrifuged Fat Graft
	Comments
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.310
	Comments
	Method	Wilcoxon (Mann-Whitney)
	Comments

2. Secondary Outcome

Title	Immunophenotyping
Description	Immunophenotyping of the fresh stromal vascular fraction of both groups. Immunophenotyping or flow cytometry measures how many cells, in a sample, express a specific surface marker. A surface marker or a group of markers may characterize a specific cell type. The software that accompanies the flow cytometer determines the number of cells (in percentage) that express the tested surface marker.
Time Frame	baseline

Outcome Measure Data

Analysis Population Description

we had technical problems in processing 4 tissue samples in each arm.

Arm/Group Title	Centrifuged Fat Graft	ADSCs Enriched Centrifuged Fat Graft
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.	female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.
Measure Participants	4	7
Mean (Standard Deviation) [percentage of expression for CD90]	27 (27.9)	79 (15.4)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Centrifuged Fat Graft, ADSCs Enriched Centrifuged Fat Graft
	Comments
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.026
	Comments
	Method	Wilcoxon (Mann-Whitney)
	Comments

3. Other Pre-specified Outcome

Title	Number of Participants Experiencing Fat Necrosis in the Postoperative Period
Description	Fat necrosis may occur whenever a fat graft is performed and it has clinical relevance. It can emerge as oil cysts or small nodules a little bit painful. In mammograms of normal breasts, fat necrosis present as cysts or micro calcifications that present a benign appearance. In breast reconstruction patients, fat necrosis, despite its benign characteristics, can suggest cancer recurrence.
Time Frame	up to 3 years

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Centrifuged Fat Graft	ADSCs Enriched Centrifuged Fat Graft
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.	female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.
Measure Participants	8	11
Number [participants]	0 0%	4 36.4%

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Centrifuged Fat Graft, ADSCs Enriched Centrifuged Fat Graft
	Comments
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.103
	Comments
	Method	Fisher Exact
	Comments

Adverse Events

Time Frame	from fat grafting to 3 years (average) for ADSC arm and 16 months for the centrifuged arm.
Arm/Group Title	Centrifuged Fat Graft		ADSCs Enriched Centrifuged Fat Graft
Arm/Group Description	female patients who underwent breast reconstruction and present with volume insufficiency will undergo centrifuged fat graft for contour and volume refinements. centrifuged fat graft: fat from the abdominal subcutaneous tissue will be taken by vacuum assisted lipectomy and immediately prepared to be grafted in the reconstructed breast that presents contour irregularities and/or volume insufficiency. No adipose derived stem cells will enrich the fat grafts in this group.		female patients who underwent breast reconstruction and present volume insufficiency will undergo ADSCs enriched fat grafting for volume and irregularity contour improvement ADSCs enriched fat graft: fat from the abdominal subcutaneous tissue will be taken by suction assisted lipectomy and stromal vascular fraction will be isolated and immediately added to the fat graft that will be employed to improve contour irregularities and volume insufficiency of reconstructed breasts.
All Cause Mortality
	Centrifuged Fat Graft		ADSCs Enriched Centrifuged Fat Graft
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	/ (NaN)		/ (NaN)
Serious Adverse Events
	Centrifuged Fat Graft		ADSCs Enriched Centrifuged Fat Graft
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/8 (0%)		0/11 (0%)
Other (Not Including Serious) Adverse Events
	Centrifuged Fat Graft		ADSCs Enriched Centrifuged Fat Graft
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/8 (0%)		0/11 (0%)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.