New and Emerging Strategies in Platelet-Rich Plasma Application in Musculoskeletal Regenerative Procedures: General Overview on Still Open Questions and Outlook

Table 4

In vivo studies on musculoskeletal tissue regeneration.

Tissue type	PRP formulation	PRP; combination	Blood volume (mL)	Centrifugation; (number of spins)	PRP volume; (mL)	Platelet concentration	Activator; (+/−)	Leukocytes; (+/−)	PAW classification	Reference
Mice dorsum	PRP	5–10–15% PRP + ADSCs-microspheres	30 mL	2	NS	NS	500 U bovine thrombin in 1 mL 10% CaCl2	NS	x	[12]
Rat limb	PRP	L-M-H PRP + BMSCs	40 mL	2	NS	High: 4358 ± 265 × 10^3; Medium: 1453 ± 88 × 10^3; Low: 48 ± 29 × 10^3; (platelets/μL)	CaCl2/thrombin	NS	P4-x; P4-x; P1-x	[13]
Sheep; sternum	PRGF	PRGF	40 mL	1	NS	NS	CaCl2	NS	x	[30]
Rat; tibia	PRP	PRP	20 mL	2	10–15% of total	1 × 10^6/μL	CaCl2	NS	P3-x	[31]
Rat; calvaria	PRP	PRP	3.15 mL	2	0.35 mL	1852.307 ± 1084.85 × 10^3/µL	CaCl2	NS	P4-x	[32]
Rat; calvaria	PRP	PRP	3.15 mL	2	0.35 mL	2977.66 ± 1174.83 × 10^3/; µL	10% CaCl2; 25% thromboplastin	NS	P4-x	[33]
Rat; femur	PRP	PRP	NS	2	NS	NS	CaCl2/; thrombin	NS	x	[34]
Mouse; calvaria	PRF	PRF	NS	NS	NS	NS	NS	NS	Not applicable	[35]
Rabbit; calvaria	PRP	PRP	10 mL	1	~0.9 mL	NS	CaCl2	NS	x	[36]
Rabbit; calvaria	PRP; PPP	PRP; PPP	130 mL	2	NS	900,000/µL	Thrombin/; CaCl2	NS	P3-x	[37]
Ovx. rat; femur	PRP	-H-PRP; -M-PRP; -L-PRP; -PPP	NS	3	NS	H-PRP: 8.21 ± 0.4 × 10^9/mL; M-PRP: 2.65 ± 0.2 × 10^9/mL; L-PRP: 0.85 ± 0.16 × 10^9/mL; PPP: 8 ± 0.5 × 10^6/mL	Thrombin/; CaCl2	NS	P4-x; P4-x; P3-x; P3-x	[38]
Ovx. mice; femur	PRP	PRP	NS	NS	NS	NS	NS	NS	Not applicable	[39]
Mini. pig; tibia	PRP	PRP + AB	120 mL	1	20 mL	NS	Thrombin/; CaCl2	NS	x	[40]
Rabbit; tibia	PRP	PRP + AB	10 mL	2	NS	NS	CaCl2/; thrombin	NS	x	[41]
Rabbit; tibia	PRP	PRP + AB	10 mL	1	0.5 mL	NS	CaCl2	NS	x	[42]
Rat; calvaria	PRP	PRP 50–100–150 µL + AB	3.15 mL	2	0.35 mL	2611.80 ±; 313.34 × 10^3/µL	CaCl2	NS	P4-x	[43]
Rabbit; calvaria	PRP	PRP + AB	40 mL	1	NS	NS	CaCl2	NS	x	[44]
Rat; calvaria	PRP	PRP 50–100–150 µL + AB	3.15 mL	2	0.35 mL	2,718.46 ± 359.65 × 10^3 platelets/µL.	CaCl2	NS	P4-x	[45]
Rabbit; femur	PRP	PRP + Bioss Collagen	4 mL	NS	NS	NS	NS	NS	Not applicable	[46]
Dog tibia	PRP	PRP + Bio-Oss	20 mL	2	NS	1.380.000	CaCl2/; thrombin	NS	Not applicable	[47]
Rat; calvaria	PRP	PRP + FFBA	3.15 mL	2	0.35 mL	2,628.8 ± 603.07 × 10^3/µL	CaCl2	NS	P4-x	[48]
Rabbit; calvaria	PRP	PRP + particulated AB	38 mL	1	5 mL	1,7 × 10^6  ±  680,200/µL	Thrombin/; CaCl2	13.42 × 10^9/L	P4-x	[49]
Rabbit; calvaria	PRP	PRP + AB	15 mL	2	1 mL	2,4 × 10^6  ±; 1,5 × 10^6/μL	CaCl2	NS	P4-x	[50]
Rabbit; calvaria	PRP	PRP + AB	15 mL	2	1 mL	2.564 × 10^6  ±; 1.621 × 10^6/μL	CaCl2	NS	P4-x	[51]
Rabbit; calvaria	PRP	PRP^* + AB; PRP° + AB	10 mL	1^*; 2°	1 mL^*; 0.5 mL°	NS	CaCl2	NS	x	[52]
Rabbit; calvaria	PRP	hPRP + AB	10 mL	2	1 mL	2.414.720 ± 1547.862 platelets/mL	CaCl2/; thrombin	NS	P1-x	[53]
Rabbit; calvaria	PRP	PRP + AB + receptor bed	10 mL	2	NS	1,2 × 10^6 platelets/mm^3	Thrombin/; CaCl2	NS	P3-x	[54]
Goat’s; frontal bone	PRP	PRP + AB	250 mL	NS	NS	800 × 10^9/L	CaCl2/; bovine thrombin	NS	P3-x	[55]
Goat’s; frontal bone	PRP	PRP + AB + BioOss	250 cm^3	4	NS	800–1000 × 10^9/L	CaCl2/; bovine thrombin	NS	P3-x	[56]
Rabbit; femur	PRGF	PRGF + DBBM	10 mL	1	1 mL	NS	CaCl2	NS	x	[57]
Rabbit; radius	PRP	hPRP + HA	500 mL	1	NS	2,422 × 10^9L	NS	NS	P4	[58]
Mini. pig’s; subcutis	PRP	PRP + Bovine HA or Phycogenic HA or Bioglass	250 mL	NS	NS	483.8 ± 97.2 × 10^3/µL	NS	24.8 ± 8.9 × 10^3/µL	P2-A	[59]
Rabbit; condyle	PRP	PRP + CPC	NS	NS	NS	NS	NS	NS	Not applicable	[60]
Mini. pig; tibia	PRP	PRP + Ca-P	120 mL	2	20 mL	NS	Thrombin/; CaCl2	NS	x	[61]
Rat; cranium	PRP	PRP + HA or β-TCP	NS	NS	NS	NS	CaCl2/; thrombin	NS	x	[62]
Rabbit; radius	PRP	PRP + BG	10 mL	1	0.8 mL	NS	CaCl2/; thrombin	NS	x	[63]
Rabbit; calvaria	PRP	PRP + rhBMP2-BCP	10 mL	2	NS	78.1 ± 1.62 × 10^4/mm^3	CaCl2/; thrombin	NS	P3-x	[64]
Rabbit; iliac crest	PRP	PRP + ringed PTFE vascular grafts + HA or collagen gel beads	5 mL	2	0.18 mL	1,100 ± 332 × 10^3/µL	NS	NS	P3	[65]
Rabbit; radius	PRP	PRP + Ti + Bone	10 mL	2	1 mL	1.0–; 1.5 × 10^12 thrombocytes/L	Thrombin/; CaCl2	NS	P3-x	[66]
Nude rat; calvaria	PRP	PRP + PGA + rhBMP-2	NS	NS	NS	NS	CaCl2/; thrombin	NS	x	[67]
Rat; femur	PRP	PRP + PCL-TCP	10 mL	2	1.5 mL	~600 × 10^3platelets/mL	CaCl2/; thrombin	NS	P2-x	[68]
Rat; ulna	PRP	PRP + gelatin hydrogel + SEW2871-micelles	10 mL	2	1 mL	NS	CaCl2	NS	x	[69]
Rabbit; calvaria	PRP	Activated/inactivated PRP + gelatine hydrogel	NS	NS	~0.8 mL	NS	Thrombin/; CaCl2	NS	x	[70]
Cattle; hoof	PRP	PRP + Gelatin microspheres + alginate	NS	2	1 mL	152.8 ± 98.7 × 10^4 platelets/μL	CaCl2	NS	P4-x	[71]
Rabbit; radius	PRP	hPRP + Persian Gulf Coral	500 mL	NS	NS	2422 × 10^9/L	NS	NS	P4	[72]
Rabbit; radius	PRP	hPRP + coral	NS	NS	NS	2422 × 10^9/L	NS	NS	P4	[73]
Rabbit; cranium	PRP	PRP + chitosan sponge	8 mL	2	0.6-0.7 mL	NS	CaCl2	NS	x	[74]
Rabbit; fibula	PRP	APC + HBO	2.5 mL	1	1.5 mL	NS	NS	NS	Not applicable	[75]
Beagles; calvaria	PRP	PRP + PDBM or Lactosorb or BMP	27 mL	1	3 mL	NS	CaCl2/; thrombin	NS	x	[76]
Goats; cranium	PRP	rPRP or hPRP or gPRP + HA or β-TCP; hPRP + bone	500 mL^h; 250 cm^3 g	NS	NS	NS	CaCl2/; thrombin	NS	x	[77]
Rabbit; calvaria	PRP	PRP + β-TCP	8 mL	2	0.7 mL	NS	NS	NS	Not applicable	[78]
Rabbit; spinal fusion	PRP	PRP + Calcium Carbonate + HA	54 mL	2	10 mL	NS	thrombin/; CaCl2	NS	x	[79]
Ovx. rat; calvaria	PRP	PRP + CA + Mel	4 mL	NS	325 µL	NS	NS	NS	Not applicable	[80]
Porcine skull	PRP	PRP + ankyloss graft	NS	NS	NS	118 ± 12; thrombocytes 1000/µL	CaCl2/; thrombin	4.3 ± 1.9 leukocytes; 1000/µL	P1-x-B	[81]
Goats tibia	PRP	PRP liquid or PRP gel + non-coated or Ca-P-coated implant	250 mL	NS	7.5 mL	800–1200 × 10^6/mL	CaCl2/; bovine thrombin	NS	P3-x	[82]
Rabbit; calvaria	PRP	PRP + BMSCs	10 mL	2	1 mL	NS	NS	NS	Not applicable	[83]
Rabbit; calvaria	PRP	PRP + induced/uninduced BMSCs	7 mL	2	NS	NS	CaCl2/; thrombin	NS	x	[84]
Mice; calvaria	PRP	PRP + BMSCs	2.0 mL	2	NS	NS	10% calcium gluconate	NS	x	[85]
Sheep; tibia	PRP	hPRP + ADSCs		2	NS	1.0 × 10^9/mL	CaCl2/; thrombin	NS	P3-x	[86]
Nude mice; inguinal groove	PRP	hPRP + hADSCs	400 mL	3	NS	NS	Thrombin/; CaCl2	NS	x	[87]
Rat calvaria	PRP	PRP + BCP or MSCs	NS	2	0.5 mL	NS	NS	NS	Not applicable	[88]
Rabbit; radius	PRP	PRP + CDHA + BMSC	NS	NS	NS	NS	CaCl2/; thrombin	NS	x	[89]
Rabbit; radius	PRP	PRP + CDHA + MSC	NS	NS	NS	NS	Thrombin/; CaCl2	NS	x	[90]
Rabbit; ulna	PRP Gel Membrane	PRP + rBMSCs + nanohydroxypaptite poly (ester urethane)	NS	3	NS	3 × 10^6 platelets/µL	NS	NS	P4	[91]
Rabbit tibia	PRP	PRP + β-TCP + BMC	9 mL	1	NS	NS	Calcium gluconate	NS	x	[92]
Nude mice cranium	PRP	PRP + β-TCP +; BMAC	27 mL	NS	NS	NS	NS	NS	Not applicable	[93]
Rabbit; calvaria	PRPGF	PRGF + Nano-HA granule + MSCs	10 mL	1	1 mL	NS	CaCl2	NS	x	[94]
Rabbit; radius	PRP	PRP + DPB + MSCs	60 mL	2	NS	12.87 ± 0.848 ×; 10^6/μL	CaCl2/; thrombin	NS	P4-x	[95]
Rat; calvaria	PRP	PRP + BiOss + BMSCs	3 mL	2	0.5 mL	NS	NS	NS	Not applicable	[18]
Rabbit patellar tendon	PRP	PRP	8 mL	1	2 mL	NS	NS	NS	Not applicable	[96]
Rabbit patellar tendon	PRP	PRP	8 mL	1	2 mL	NS	NS	NS	Not applicable	[97]
Rabbit Achilles tendon	PRP	PRP	8 mL	1	2 mL	NS	NS	NS	Not applicable	[98]
Horse SDFT	PRP	PRP	NS	NS	3 mL	NS	NS	NS	Not applicable	[99]
Horse SDFT	PRP	PRP	NS	NS	3 mL	639.7 ± 103.2 × 10^9/L	NS	42.1 ± 16.7 × 10^9/L	P2-A	[100]
Rat patellar tendons	PRP	PRP	60 mL	2	NS	523.8 × 10^4/mL	NS	NS	P1	[101]
Rat; rotator cuff	PRP	PRP	5 mL	2	NS	13.8 × 10^9 platelets/L	NS	NS	P1	[102]
Rat Achilles tendon	PRP	PRP	NS	2	NS	(2.2–2.9) × 10^6/mm^3	CaCl2	1 × 10^2/mm^3	P4-x-B	[103]
Rat; tendon-from-bone supraspinatus tear	PRP	PRP	7-8 mL	2	NS	NS	CaCl2/thrombin	NS	x	[104]
Rabbit; Achilles tendon	PRP	PRP	8 mL	NS	2 mL	NS	NS	NS	Not applicable	[105]
Sheep Achilles tendon	PRGF	PRGF	20 mL	1	2 mL	NS	CaCl2	NS	x	[106]
Rabbit; intrasynovial flexor tendons	PRPF	PRP/PRPF	10 mL	2	ns	313.5 (±72.3) × 10^4 μL	CaCl2/thrombin	NS	P4-x	[107]
Rat patellar tendon	PC	PC	9 mL	1	0.5 mL	445–862 thousand/mm^3	NS	NS	P2/P3-x	[108]
Sheep; DDFT	PRP	PRP + 10 × 10^6 autologous PB-MSCs	18 mL	NS	3/5 mL	882 ± 199 × 10^3 platelet/μL	NS	NS	P3	[109]
Rabbit; Achilles tendon	PRP	PRP + 1 × 10^7 ADSCs	10 mL	2	NS	NS	CaCl2	NS	x	[110]
Sheep Achilles tendon	PRPFM	PRPFM + APD	9 mL	NS	NS	NS	NS	NS	Not applicable	[111]
Rat Achilles tendon	PRP	PRP + Collagen sponge + TSCs	20 mL	2	NS	8 × 10^9 platelets/L	NS	NS	P1	[112]
Mice abdominal cavities	PRCR	10% PRCR in DCs	50 mL	2	NS	10.0 × 10^8 platelets/mL	CaCl2	NS	P3-x	[113]
Rabbit articular cartilage	PRP	PRP + PLGA	16 mL	2	0.8 mL	125.59 × 10^4/μL	NS	NS	P4	[114]
Sheep articular cartilage	PRP	PRP + collagen-HA	20 mL	1	NS	874 ± 87 × 10^3/μL	CaCl2	NS	P3-x	[115]
Caprine articular cartilage	PRP	PRP + DBM	27 mL	1	3 mL	1511 × 10^9/L	NS	NS	P4	[116]
Rabbit articular cartilage	PRP	PRP + chondrocytes	10 mL	2	1 mL	NS	NS	NS	Not applicable	[117]
Rabbit articular cartilage	PRP	PRP + BMSCs; PRP + ADSCs	18 mL	2	0.5 mL	16.7 ± 1.1 × 10^8/mL	Ca	NS	P4-x	[118]
Rabbit articular cartilage	PRP	PRP + SDMC	9 mL	2	1 mL	2103.56 ± 479.5 × 10^3/mL	NS	NS	P4	[119]
Rat articular cartilage	PRP	PRP + hydrogel + chondrocytes	40 mL	2	NS	1240.9 ± 86.1 × 10^4/µL	NS	NS	P4	[120]
Rabbit articular cartilage	PRP	PRP + hydrogel + chondrocytes	10 mL	2	NS	NS	NS	NS	Not applicable	[121]
Mini pig articular cartilage	PRP	PRP + BMSCs + biphasic scaffold	60 mL	1	10 mL	2.2 ± 0.57 × 10^6 cells	Thrombin/CaCl2	NS	Not applicable	[122]
Rabbit articular cartilage	PRP	Autologous cartilage fragments + hyaluronic acid derivative + human fibrin glue + PRP	8 mL	2	400 µL	858 × 10^3 platelets/mm^3 (range 642–1,085)	NS	NS	P3	[123]
Sheep cartilage	PRP	PRP + microfractures	60 mL	2	6–8 mL	1415 ± 164 × 10^3/mL	Fibrinogen; Thrombin	NS	P1-x	[124]
Rat articular cartilage	PRP	PRP + microfractures	NS	2	NS	13.8 × 10^9 platelets/L	NS	NS	P1	[125]
Porcine ACL	PRP	Collagen-PRP hydrogel	60 mL	2	9 mL	NS	NS	NS	Not applicable	[126]
Canine ACL	PRP	Collagen-PRP hydrogel	NS	1	NS	NS	NS	NS	Not applicable	[127]
Canine ACL	PRP	Collagen-PRP hydrogel	NS	NS	NS	NS	NS	+	Not applicable	[128]
Mini. pig ACL	PRP	3X or 5X PRP + collagen scaffold	NS	NS	NS	1951 ± 304 × 10^9/L; 1161 ± 179 × 10^9/L	NS	NS	P4; P3	[129]
Rabbit; ACL	PRP	PRP + DPB	NS	2	NS	NS	CaCl2/bovine thrombin	NS	x	[130]
Canine ACL	PRP	PRP	20 mL	3	1 mL	669 ± 51 × 10^9 platelets/L	NS	NS	P2	[131]

Not specified (NS), biphasic calcium phosphate (BCP), mesenchymal stem cell (MSC), beta-tricalcium phosphate (β-TCP), polytetrafluoroethylene (PTFE), platelet-rich growth factor (PRGF), autologous bone (AB), hydroxyapatite (HA), calcium phosphate cement (CPC), bone marrow mesenchymal stem cells (BMSCs), calcium aluminate (CA), rabbit BMSC (rBMSC), deproteinized bovine bone mineral (DBBM), calcium-deficient hydroxyapatite (CDHA), allogeneic-mesenchymal stem cells (aMSC), titanium (Ti), human adipose derived stem cell (hADSC), human PRP (hPRP), fresh frozen bone allograft (FFBA), autogenous bone (AB), hyperbaric oxygen (HBO) therapy, autologous platelet concentrate (APC), adipose tissue derived stem cells (ASC), calcium phosphate (Ca-P), autogenous particled bone (APB), allogeneic BMSC (aBMSCs), acellular porcine dermal patch (APD) polyglycolic acid (PGA), recombinant human bone morphogenetic proteins (rhBMP2), platelet-poor plasma (PPP), rat PRP (rPRP), human PRP (hPRP), goat (gPRP), perforated bone matrix (PDBM), bone morphogenetic proteins (BMP), polycaprolactone-20% tricalcium phosphate (PCL-TCP), (5-[4-phenyl-5-(trifluoromethyl) thiophen-2-yl]-3-[3-(trifluoromethyl) phenyl] 1,2,4-oxadiazole) SEW2871, borate glass (BG), platelet-rich fibrin (PRF), deproteinized bone matrix (DPB), low-medium-high PRP (L-M-H PRP), bone marrow cells (BMC), adipose derived stem cell (ADSC) bone marrow aspirate concentrate (BMAC), superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFT), anterior cruciate ligament (ACL), platelet-rich plasma fibrin matrix (PRPFM), concentrate plasma (PC), platelet-rich plasma-clot release (PRCR), plasma rich in growth factors (PRGF), PRP with fibrin matrix (PRPF), tendon stem cells (TSCs), periferal blood mesenchymal stem cells (PB-MSCs), adipose derived mesenchymal stem cells (ADSCs), white blood cells (WBC), poly(lactic-co-glycolic acid) (PLGA), demineralized bone matrix (DBM), hydroxyapatite (HA), synovial membrane derived mesenchymal stem cell (SDMC), bone marrow mesenchymal stem cell (BMSC), adipose derived stem cell (ADSC), deproteinized bone (DPB), and calcium chloride (CaCl2), calcium (Ca).
^*Anitua method. °Sonnleitner method.
PAW classification: P1 (≤baseline), P2 (>baseline-750,000), P3 (>750,000–1,250,000), P4 (>1,250,000), x (exogenous activation), A (above baseline), B (below baseline), h (human), and g (goat).