In vitro: hSVF () or hADSCs () from subcutis of 3 donors (38-52 yrs) THP1 () In vivo: 46 nude rats with bilateral bone defects () in femoral condyles hSVF () or hADSCs () from abdomen THP1 ()
In vitro: Group 1: THP1 Group 2: SVF Group 3: THP1+SVF Group 4: ADSCs Group 5: THP1+ADSCs In vivo: Group 1: no treatment Group 2: THP1 Group 3: SVF Group 4: THP1+SVF Group 5: ADSCs Group 6): THP1+ADSCs
In vitro: ALP activity Calcium content In vivo: Histology Histomorphometry
In vitro: 2 wks Group 3: ↑ ALP activity than groups 2, 4, and 5. 1 mo Group 3: ↑ calcium content than groups 2 and 5 In vivo: 4 and 10 wks Groups 3, 4, 5, and 6: ↑ BA/TA than groups 1 and 2. 10 wks Groups 3 and 4: ↑ BA/TA than groups 5 and 6
In vitro: hSVF () from 8 donors () PLCL scaffold COLL I/COLL III scaffold
Group 1: PLCL+SVF in NM Group 2: COLL I/COLL III+SVF in NM Group 3: PLCL+SVF in CM Group 4: PLCL+SVF in OM Group 5: COLL I/COLL III+SVF in CM Group 6: COLL I/COLL III+SVF in OM
RT-PCR (ACAN, SOX6, SOX9, ALP, ONC, COLL I, COLL III, and COLL X)
4 days Group 3: ↑ ACAN gene expression than group 5) 2 wks Groups 3 and 5: ↑ SOX6, SOX9 gene expression during time Groups 4 and 6: ↑ ALP, ONC, and COLL I gene expression during time 3 wks Group 5: ↑ COLL II gene expression than group 3 Groups 3 and 5: ↓ COLL X gene expression during time
In vitro: hSVF or hADSCs () from 7 donors Xenohybrid bone graft scaffold ()
Group 1: SVF in plastic in NM Group 2: ADSCs in plastic in NM Group 3: scaffold+SVF in NM Group 4: scaffold+ADSC in NM Group 5: SVF in plastic in OM Group 6: ADSCs in plastic in OM Group 7: scaffold+SVF in OM Group 8: scaffold+ADSC in OM
Micro-CT IHC (OCN) ALP activity Mineralization
2 mo Group 7: ↑ bone trabeculae than groups 4, 8, and 3 Groups 3 and 4: ↓ OCN than groups 7 and 8 Group 1: ↑ ALP activity than group 2 Groups 1 and 2: ↓ mineralization than groups 5 and 6
In vivo: CD1nu/nu athymic mice with subcutaneous pouches hSVF from abdomen or breast of donors () βTCP scaffold (8 mm diameter, 4 mm height) HA scaffold (8 mm diameter, 4 mm height)
Group 1: βTCP+SVF ( CFU-f/cm3) Group 2: βTCP+SVF ( CFU-f/cm3) Group 3: βTCP+SVF ( CFU-f/cm3) Group 4: HA+SVF ( CFU-f/cm3) Group 5: HA+SVF ( CFU-f/cm3) Group 6: HA+SVF ( CFU-f/cm3)
Histology
2 mo Groups 1, 4, 3, and 6: ↑ dense matrix than groups 2 and 5 Groups 3 and 6: ↑ dense matrix similar to osteoid than groups 1 and 4
In vivo: Nude mice with subcutaneous pouches Nude rats with bilateral defects in parietal bone (4 mm) Devitalized hypertrophic cartilage pellet from 5 donors () hSVF from 12 donors ()
Group 1: pellet in pouches Group 2: pellet+SVF (/ml) in pouches Group 3: pellet+SVF (/ml) in pouches Group 4: pellet+SVF (/ml) in pouches Group 5: pellet in bone defects Group 6: pellet+SVF (/ml) in bone defects
Micro-CT Histology
3 mo Group 3: ↑ mineralized volume, BA than group 1 Groups 2 and 3: ↑ vessel length density than group 1 1 mo Group 6: ↑ BA than group 5
In vivo: 24 syngenic Balb/c mice (8 wks) with subcutaneous pouches Autologous SVF () from epididymis Bio-Oss® scaffold (deproteinized sterilized bovine bone) (10 mg)
Group 1: scaffold Group 2: scaffold+SVF+PRP
Histology IHC (OPN) Histomorphometry RT-PCR (OSX, OCN, COLL I, and ALP)
2 wks Group 2: ↑ OPN protein than group 1 2 and 4 wks Group 2: ↑ OSX, OCN gene expression than group 1 1, 2, and 4 wks Group 2: ↑ COLL I gene expression than group 1 2 mo Group 2: ↑ ALP gene expression, osteoid-like tissue, OPN protein, vascularization than group 1
In vivo: 14 nude immunocompromised mice (8 wks) with bilateral critically sized calvarial defects Nude mice with muscular pouches hSVF or hADSCs () from flank, scapula, abdomen, or inner thigh of donors PCL-DCB scaffold (4 mm diameter, 0.64 mm length)
Group 1: scaffold in bone defect Group 2: scaffold+SVF in bone defect Group 3: scaffold+ADSCs in bone defect Group 4: scaffold in pouches Group 5: scaffold+SVF in pouches Group 6: scaffold+ADSCs in pouches
Micro-CT Histology
3 mo Groups 2 and 3: ↑ BV than group 1 Group 2: ↑ BV during time 10 days Group 5: ↑ VA than groups 4 and 6 6 wks Groups 5 and 6: ↑ VA than group 4
In vivo: 50 SD rats (8 wks) with critically sized calvarial defects Autologous SVF () from inguinal region DBM scaffold PLA scaffold
Group 1: no treatment Group 2: DBM Group 3: DBM+SVF Group 4: DBM+PLA Group 5: DBM+PLA+SVF
Gross evaluations Histology IHC (OCN)
2 mo Groups 3 and 5: ↑ defect filling, BA, OCN than groups 1, 2, and 4 Group 2: ↑ defect filling than groups 1 and 4 Group 4: ↑ defect filling than group 1
In vivo: 15 NZW rabbits (14-16 wks) with unilateral segmental bone defect in the middiaphysis of left ulna (20 mm) Allogenic SVF () from suprascapular region PLGA scaffold ()
Group 1: scaffold Group 2: scaffold+SVF Group 3: scaffold+osteogenically differentiated SVF
Radiography Micro-CT Histology
2 mo Group 3: defect bridging, trabecular bone with smooth margins Group 3: ↑ BVF than groups 1 and 2 Group 3: ↑ Tb.Th than group 1 Groups 2 and 3: scaffold absorbed Group 2: ↑ BVF than group 1
Clinical trial: 8 pz (62-84 yrs) with displaced low-energy fractures of the proximal humerus Autologous SVF from abdomen Fibrin hydrogel+porous silicate HA microgranules scaffold In vivo: Adult nude rats with a critically sized segmental defect in femora (6 mm) Fibrin hydrogel+porous silicate HA microgranules scaffold hSVF () from abdomen
Clinical trial: Group 1: scaffold+SVF pelleted In vivo: Group 1: scaffold Group 2: scaffold+SVF
Clinical trial: Safety Histology Micro-CT In vivo: Micro-CT Histology
Clinical trial: 12 mo Group 1: no adverse reactions; ↓ pain during time; ↑ formed bone ossicles during time In vivo: 2 mo Group 2: ↑ mineralized volume, BV/TV, newly formed bone, frank bone, early stage of maturation than group 1
