Personalized Cell Therapy: scientific background and mechanism of

action
Iva Borovečki Žuti, dr. med.
PRP – BACKGROUND
Properties of PRP

PLATELETS AND THEIR ACTIVATION

(Thrombin)
(CaCl2)
Collagen

▪ Dense bodies ▪ Release of > 300 GF and

• Serotonin, ADP, Calcium cytokines
▪ α-Granula
• Growth factors (GF)
Growth Factors in Platelets

FGF
PDGF
(Fibroblast Growth Factor)
(Platelet Derived Growth Factor)
Growth and differentiation of
Cell growth, chemotaxis,
chondrocytes and osteoblasts,
collagen production
collagen production

EGF TGF-ß
(Epidermal Growth Factor) (Transforming Growth Factor)
Platelets
Endothelial chemotaxis / Proliferation, mitogenesis,
angiogenesis, epithelial collagen synthesis, regulates
mitogenesis macrophage and lymphocyte
proliferation

VEGF
(Vascular endothelial Growth Factor)
Angiogenesis, mitogenesis of
endothelial cells
Properties of PRP:

GROWTH FACTORS
The growth factors of the PRP mediate
numerous processes:
▪ Proliferation of cells
▪ Chemotaxis
▪ Differentiation of cells
▪ Angiogenesis

Collagen synthesis, formation of ECM,

tissue regeneration

Arshdeep M.; PRP in dermatology: Boon or a bane? IJDVL 2014; 80

(1): 5-14
ACP – Autologous Conditioned Plasma

ACP

A C P
Autologous ➔ Conditioned ➔ separated Plasma ➔ blood plasma with
Body’s own and concentrated its components

36
ACP – The Healing Composition

Platelet concentration
600

10^3/µl
400 2-3x
200
Plasma with
Platelets 0
Centrifugation Blood ACP

RBCs, WBC concentration

WBCs
6

10^3/µl
Blood ACP 4 -80%
2

0
Blood ACP

42
ACP – completely autologous

1,500 rpm / Closed System

5 min Transfer

Aspiration of Ready for

15cc Blood Injection

41
PRP Requirements

▪ Increased concentration of PLATELETS (>2x whole blood)

• Proven significant proliferation stimulation and hyaluronic acid synthesis upon platelet
concentrations >2x whole blood1,2

1 Mazzocca et al. The positive effects of different platelet-rich plasma methods on human muscle, bone, and tendon cells. AJSM 2012;40: 1742-9
2 Anitua et al. Fibroblastic response to treatment with different preparations rich in growth factors. Cell Prolif. 2009; 42:162-170
10
Adipose-derived mesenchymal stem cells

SVF
▪ Stromal vascular fraction
▪ Source of adipose-derived stem
cells and growth factors
• Reduction of local
inflammation1
• Prevention of tissue fibrosis1
• Anti-apoptotic2
• Supports angiogenesis and
tissue remodeling2

• 1 Luz-Crawford et al, Mesenchymal stem cell derived IL1RA promotes macrophage polarization and inhibits B cell differentiation. Stem Cells 2016;34:483–92.
• 2 Pers et al, Adipose Mesenchymal Stromal Cell-Based Therapy for Severe Osteoarthritis of the Knee: A Phase 1 Dose-Escalation Trial. 2016; 5:847-856
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ACASVF – AdSC concentration

AdSCs/cc
Oil 1,400,000

Processing and 1,200,000 4-5x

centrifugation
ACASV 1,000,000

F 800,000

Fat ACAS 600,000

graft VF
400,000

Adipocytes Pre-adipocytes 200,000

Pericytes Endothelial 0
and progenitor cells Fat graft ACASVF
ACASVF
Extracellular
Adipose- Hematopoetic cells matrix
derived stem cells Monocytes / macrophages Data on file.

• 30 ml of lipoaspirate=1.5 ml of SVF
• 3.5x10^5 per 1 ml viable nucleated cells of lipoaspirate (dry, hence, removal of any oil and
tumescent solution)
• SVF it is ~2x10^6 NCL per ml of SVF
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Preparation of ACASVF

Fat tissue harvesting from an appropriate 1st Centrifugation at 2500rpm for 4min. Remove the syringe, avoid mixing.
donor site.
Please follow appropriate fat tissue
harvesting protocol.

30x

Isolation of fat graft. Processing via 1.4mm transfer 2nd centrifugation at 2500rpm Isolation of SVF
device. for 4min. ACASVF
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ACPSVF – the regenerative match

Principle of ACPSVF Benefits:

▪ Source of growth factors
ACP ACPSVF ▪ Source of regenerative cells
Growth factors Adipose-derived
stem cells (AdSC) ▪ Growth and proliferation stimulating effect of ACP
on AdSCs shown in vitro1

ACPSVF
Regenerative
Preparation
match
ACP 5x

SVF

1 Loibl et al, The effect of leukocyte-reduced platelet-rich plasma on the proliferation of autologous adipose-tissue derived mesenchymal stem cells. Clin Hemorheol Microcirc, 2016;61(4):599-614
PRP – INDICATIONS
PRP – Aesthetic Medicine

SKIN REJUVENATION:
▪ PRP stimulates the removal of photodamaged extracellular
matrix1
▪ PRP induces the synthesis of collagen1
▪ PRP stimulates the synthesis of the body’s own hyaluronic
acid2

Filling of volume deficit, skin tightening and thickening of the

epidermis
PRP – Aesthetic Medicine

SKIN REJUVENATION – UNDEREYE AREA:

▪ Indications:
• Loss of volume
• Dark circles
• Cutis laxa Aust et al.2

▪ Difficulties with other treatment options:

• Edema (HA); Ectropion1 (Botox); Blepharoplasty
(risks of operation)

1 Ozgur et al. Dry eye syndrome due to botulinum toxin type-A injection: guideline for prevention. Hawaii J Med Public Health 2012; May;71(5): 120-3
2 Aust et al. Platelet-Rich Plasma – Eine minimal-invasive Methode zur Behandlung alternder Haut am Unterlid. Face 1/2016,
PRP – Aesthetic Medicine

(ANDROGENIC) ALOPECIA:
▪ PRP stimulates the proliferation and
differentiation of stem cells in the hair follicle
bulge area1

Increase in hair diameter and

density2-4
Kathu et al.4

1 Li et al. Autologous platelet-rich plasma: A potential therapeutic tool for promoting hair growth. Dermatol Surg 2012;38:1040-6
2 Greco, Brandt. The effects of autologous platelet rich plasma and various growth factors on non-transplanted miniaturized hair. Hair Transplant Forum 2009;19:49-50
3 Lopez et al. Autologous platelet-rich plasma as a potential therapeutic tool in androgenetic alopecia. J Am Acad Dermatol 2013;68:SAB 103
4 Khatu et al. Platelet-Rich Plasma in Androgenic Alopecia: Myth or an Effective Tool. J Cutan Aesthet Surg 2014;7(2):107-110
PRP – Aesthetic Medicine

COMBINATION TREATMENT
WITH MEDICAL NEEDLING
▪ Better cosmetic outcome1
▪ Higher patient satisfaction1

Sasaki et al.1

PRP + medical needling

Good combination for skin rejuvenation1

1 Sasaki. Micro-Needling Depth Penetration, Presence of Pigment Particles, and Fluorescein-Stained Platelets: Clinical Usage for Aesthetic Concerns.Aesthet Surg J. 2016 Aug 16.
PRP – Aesthetic Medicine

SUPPORTIVE TREATMENT
FOR LASER TREATMENTS
▪ PRP has proliferation-enhancing effects1
▪ PRP reduces downtime2,3
▪ Better cosmetic result1
▪ Higher patient satisfaction1

PRP + fractionated laser is a good combination Shin et al.

for skin rejuvenation1,2

1 Gawdat et al. Autologous platelet rich plasma: topical versus intradermal after fractional ablative carbon dioxide laser treatment of atrophic acne scars.Dermatol Surg. 2014
Feb;40(2):152-61
2 Na et al. Rapid healing and reduced erythema after ablative fractional carbon dioxide laser resurfacing combined with the application of autologous platelet-rich plasma. Dermatol Surg.
2011 Apr;37(4):463-8
3 Lee JW. The efficacy of autologous platelet rich plasma combined with ablative carbon dioxide fractional resurfacing for acne scars: a simultaneous split-face trial. Dermatol Surg. 2011
Jul;37(7):931-8
PRP – Plastic Surgery

RECONSTRUCTIONS:

▪ PRP stimulates the proliferation of cells1

▪ PRP supports the vascularization of fat grafts

▪ Supports the ingrowth and survival of fat grafts

and the preservation of volume

1 Loibl et al. The effect of leukocyte-reduced platelet-rich plasma on the proliferation of autologous adipose-tissue derived mesenchymal stem cells. Clin Hemorhoil Microcirc 2014;
Epub ahead of print
2 Oh et al. Activated plateled-rich plasma improves fat graft survival in a nude mice: A pilot study. Dermatol Surg 2011;37:619-25
3 Gentile et al. A Comparative Translational Study: The Combined Use of Enhanced Stromal Vascular Fraction and Platelet-Rich Plasma Improves Fat Grafting Maintenance in Breast
Reconstruction. Stem Cells Transl Med. 2012;1(4):341–351
PRP – Plastic Surgery

WOUNDS:

▪ PRP induces the upregulation of cell cycle regulatory

proteins1
▪ Antibacterial effect of PRP2,3

Accelerates the re-epithelialization process1,4 Mehrannia et al.5

1Kim et al. Application of PRP accelerates the wound healing process in acute and chronic ulcers through rapid migration and upregulation of cyclin A and CDK4 in HaCaT cells. Mol Med
Rep 2013;7:476-80
2 Moojen et al. Antimicrobial Activity of Platelet-Leukocyte Gel against Staphylococcus aureus. J Orthop Res 2007;25:1-7
3 Trowbridge et al. Use of Platelet Gel and Its Effects on Infection in Cardiac Surgery. JECT 2005;37:381-386
4 Rallis et al. Amyopathic dermatomyositis-associated bilateral elbow ulcers successfully treated with autologous PRP. Int J Dermatol. 2014 Jan;53(1)
5 Mehrannia et al. Platelet rich plasma for treatment of nonhealing diabetic foot ulcers: a case report. Can J Diabetes. 2014 Feb;38(1):5-8. doi: 10.1016/j.jcjd.2013.08.271.
Application of PRP – What Needs to be Considered?

▪ PRP should not be mixed/co-injected with local anesthetics1

▪ Numbing cream is fine

▪ Increasing effect upon multiple injections2

• Skin: Recommendation: 3-5 (intradermal) injections (monthly interval)
• Scalp: Recommendation: 3-6 injections (1.5-3mm depth) (2-4 weeks interval)

▪ The use of small cannula sizes is unproblematic3

▪ Recommendation Scalp: 32G 4mm Needle
▪ Recommendation Skin: 34G / Invisible Needle | Under-eye: 27G 38mm Cannula
1 Carofino et al. Corticosteroids and Local Anesthetics Decrease Positive Effects of PRP: An In Vitro Study on Human Tendon Cells. Arthroscopy 2012; 28 (5): 711-719
2 Aust M, et al. Platelet-rich Plasma for Skin Rejuvenation and Treatment of Actinic Elastosis in the Lower Eyelid Area. Cureus 2018 10(7): e2999.
3 Bausset et al. Impact of local anaesthetics and needle calibres used for painless PRP injections on platelet functionality. Muscles, Ligaments and Tendons Journal 2014; 4 (1): 18-23
Angel System:

▪ PRP
▪ Bone Marrow Concentrate

Closed processing
Preconnected disposable processing kit
Separate, sterile compartments
Thrombinator™
Preparation of Autologous Thrombin Serum

Preparation of thrombin serum from autologous

whole blood, PRP, PPP

Easy to use, 10 - 15 min.

Thrombin serum enables clotting (gel formation)

of PRP (ACP and Angel® PRP) and BMC

Indications: Chronic wounds, Non-unions,

delayed fracture healing, bone marrow lesions
(BML)

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Overview of Arthrex PRP Systems

ACP DOUBLE SYRINGE ANGEL SYSTEM

▪ Optimized composition ▪ Maximum flexibility, indication tailored PRP

(~ 2.5x platelets, almost free formulations
of WBCs & RBCs)
▪ PRF gel in combination with thrombin
▪ Completely autologous ▪ Enables processing of large blood quantities
(no anticoagulant)
▪ Closed system
▪ Closed system
▪ Fully-automated
▪ Convenient processing, not
▪ Fast dependent on
user skills
Arthrex Support

PRP - PUBLICATION DATABASE

▪ > 80 studies in aesthetics

▪ Regular update
▪ No login required
▪ Various display options
▪ Link to PubMed