Assessment of serum levels and placental bed tissue expression of IGF-1, bFGF, and PLGF in patients with placenta previa complicated with placenta accreta spectrum disorders

Figures & data

Table

Target gene	Primer sequence
IGF-1	F 5′-ACATTGCTCTCAACATCTCCCA-3′; R 5′-TGGTGTGCATCTTCACCTTCA-3′
bFGF	F 5′-GAAGAGCGACCCTCACATCAA −3′; R5′-ACGGTTAGCACACACTCCTTT -3′
PLGF	F 5′-GCTCGTCAGAGGTGGAAGTG-3′; R 5′-CAGCAGGGAGACACAGGATG-3′
BA	F 5′-CCTTCCTGGGCATGGAGTC-3′; R 5′-TGATCTTCATTGTGCTGGGTG-3′

Abbreviations: IGF-1: insulin-like growth factor-1; bFGF: basic fibroblast growth factor; PLGF: placental growth factor; BA: β-actin; F: forward (upstream) primer; R: reverse (downstream) primer.

Table 1. Maternal and fetal demographic and clinical features/outcomes in patients with complete placenta previa with PAS disorders and without PAS disorders.

Demographic and clinical variables	PAS	Non-PAS	P-value
	N = 25 (62.5%)	N = 15 (37.5%)
Age (years)	34.92 ± 4.02	31.27 ± 4.67	0.014
<35 years	9 (36%)	11 (73.33%)	0.048
≥35 years	16 (64%)	4 (26.67%)	0.048
Weight (kg)	71.08 ± 6.52	64.31 ± 6.04	0.023
Height (cm)	161.48 ± 4.66	161.47 ± 5.25	0.993
BMI (kg/m^2)	27.56 ± 2.77	24.64 ± 1.75	0.0007
GA at admission (weeks)	34.55 ± 1.53	35.54 ± 1.56	0.055
Gravida (G)	3.96 ± 1.77	2.07 ± 0.96	0.0004
Parita (P)	1.16 ± 0.85	0.47 ± 0.52	0.007
Prior cesarean section (CS)	1.12 ± 0.78	0.2 ± 0.41	0.0001
0 CS	4 (16%)	13 (86.67%)	0.000
1 CS	16 (64%)	2 (13.33%)	0.000
≥ 2 CS	5 (20%)	0 (0%)	0.000
Prior diagnostic/curettage (D/C)	1.72 ± 1.31	0.6 ± 0.74	0.004
0 D/C	5 (20%)	8 (53.33%)	0.058
1 D/C	10 (40%)	5 (33.33%)	0.058
≥2 D/C	10 (40%)	2 (13.33%)	0.058
Conceived through IVF	3 (12%)	0 (0%)	0.279
History of smoking	5 (20%)	1 (6.67%)	0.381
Prior placenta previa	1 (4%)	0 (%)	0.625
Prior uterine adhesions	3 (12%)	1 (6.67%)	0.516
Vaginal bleeding	11 (44%)	10 (66.67%)	0.204
Cervical length (cm)	3.73 ± 0.92	3.84 ± 0.69	0.696
Placental thickness above the cervix (cm)	3.752 ± 1.78	2.713 ± 1.08	0.048
Placental length from the cervix (cm)	4.232 ± 1.96	2.987 ± 1.57	0.043
GA at the termination of pregnancy (weeks)	35.25 ± 1.33	36.17 ± 1.41	0.046
Bleeding amount during CS (ml)	1100 ± 959.06	433.33 ± 238.05	0.012
< 100 ml    (non-PPH)	15 (60%)	14 (93.33%)	0.03
≥ 100 ml (PPH)	10 (40%)	1 (6.67%)	0.03
Blood transfusion	12 (48%)	2 (13.33%)	0.04
Autotransfusion	14 (56%)	1 (6.67%)	0.002
Transfusion products
RBC (u)	1.2 ± 2.29	0.27 ± 0.70	0.134
Plasma (ml)	108 ± 293.92	0 ± 0.00	0.165
Autotransfusion (ml)	266.2 ± 310.58	33.33 ± 129.10	0.008
Duration of the surgery (minutes)	125.81 ± 71.64	73.93 ± 14.80	0.008
Post-surgery hemoglobin (g/l)	95.68 ± 13.20	101.67 ± 16.80	0.217
Post-surgery hospitalization days	7.4 ± 1.73	6.06 ± 0.80	0.008
Newborn weight (g)	2353 ± 568.06	2742 ± 381.30	0.024
Newborn APGAR score
At 0 min	7.6 ± 1.89	8.47 ± 0.64	0.095
At 5 min	8.56 ± 2.04	9.54 ± 0.52	0.079
Admission to NICU	14 (56%)	3 (20%)	0.046

Abbreviations: PAS: Placenta accreta spectrum; IVF: in-vitro fertilization; GA: gestational age; PPH: post-partum hemorrhage; RBC: red blood cells; NICU: neonatal intensive care unit.

Note: All data presented in this table were expressed as means ± SD.

Figure 1. IGF-1, bFGF, and PLGF placental bed tissue expression at 400x magnification in complete placenta previa patients without PAS disorders (control group) and with PAS disorders (case group). (A) IGF-1 expression in placental bed tissues of patients without PAS disorders (CPP). (B) IGF-1 expression in placental bed tissues of patients with PAS disorders. (C) bFGF expression in placental bed tissues of patients without PAS disorders. (D) bFGF expression in placental bed tissues of patients with PAS disorders. (E) PLGF expression in placental bed tissues of patients without PAS disorders. (F) PLGF expression in placental bed tissues of patients with PAS disorders. (M) represents the myometrium. (V) represents the villous trophoblastic cells. (Black arrows) indicate the blue-colored granules, representing the staining intensity of the nucleus. (Red arrows) indicate the brown-yellow granules, representing the positive cells.

Table 2. Serum levels and placental bed tissue expression of IGF-1, bFGF, and PLGF in patients with complete placenta previa with PAS disorders and without PAS disorders.

Variables	PAS	Non-PAS	P-value
	N = 25 (62.5%)	N = 15 (37.5%)
Serum level (ELISA)
IGF-1 (5–160 ng/mL)	22.65 ± 4.61	29.4 ± 4.43	<0.0001
bFGF (2.5–80 pg/mL)	12.38 ± 2.20	13.71 ± 1.96	0.063
PLGF (2.5–80 pg/mL)	14.39 ± 2.58	12.38 ± 2.44	0.0200
Relative mRNA expression (qRT-PCR)
IGF-1/BA   (fold-change)	3.527 ± 0.70	1.000 ± 0.64	<0.0001
bFGF/BA   (fold-change)	2.120 ± 0.36	1.000 ± 0.42	<0.0001
PLGF/BA   (fold-change)	2.817 ± 0.79	1.000 ± 0.38	<0.0001
Staining intensity (IHC)
IGF-1(fold-change)	1.070 ± 0.08	1.000 ± 0.10	0.018
bFGF (fold-change)	1.072 ± 0.10	1.000 ± 0.11	0.033
PLGF (fold-change)	1.060 ± 0.07	1.000 ± 0.10	0.026
Positive cells count/field (IHC)
IGF-1 (count/field)	177.5 ± 20.02	167.8 ± 14.57	0.110
bFGF (count/field)	186.4 ± 19.01	180.6 ± 14.51	0.317
PLGF (count/field)	172.9 ± 17.82	167.4 ± 15.57	0.335

Abbreviations: PAS: Placenta accreta spectrum; qRT-PCR: quantitative real-time polymerase chain reaction; IHC: immunohistochemistry; IGF-1: insulin-like growth factor-1; bFGF: basic fibroblast growth factor; PLGF: placental growth factor; BA: β-actin.

Note: All data presented in this table were expressed as means ± SD.

Table 3. Serum levels and placental bed tissue expression of IGF-1, bFGF, and PLGF in non-PAS, PAS grade I, PAS grade II, and PAS grade III.

Variables	Non-PAS	PAS grade I	PAS grade II	PAS grade III	P-value
	N = 15 (37.5%)	N = 7 (17.5%)	N = 12 (30%)	N = 6 (15%)
Serum level (ELISA)
IGF-1 (5–160 ng/mL)	29.43 ± 4.43	24.30 ± 4.03	21.98 ± 3.29	22.03 ± 7.31	0.0006
bFGF (2.5–80 pg/mL)	13.70 ± 1.96	13.08 ± 2.26	11.90 ± 1.98	12.52 ± 2.67	0.1927
PLGF (2.5–80 pg/mL)	12.38 ± 2.44	12.96 ± 2.74	14.97 ± 2.56	14.89 ± 2.14	0.0392
Relative mRNA expression (qRT-PCR)
IGF-1/BA (fold-change)	1.000 ± 0.64	3.194 ± 1.40	3.509 ± 0.63	3.872 ± 0.70	<0.0001
bFGF/BA (fold-change)	1.000 ± 0.42	2.240 ± 0.31	2.140 ± 0.30	1.940 ± 0.51	<0.0001
PLGF/BA (fold-change)	1.000 ± 0.38	2.784 ± 1.14	2.810 ± 0.71	2.869 ± 0.48	<0.0001
Staining intensity count (IHC)
IGF-1 (fold-change)	1.000 ± 0.10	1.059 ± 0.10	1.063 ± 0.05	1.100 ± 0.09	0.1044
bFGF (fold-change)	1.000 ± 0.11	1.084 ± 0.10	1.047 ± 0.10	1.109 ± 0.07	0.1062
PLGF (fold-change)	1.000 ± 0.10	1.047 ± 0.04	1.053 ± 0.07	1.071 ± 0.07	0.2071
Positive cells count (IHC)
IGF-1 (count/field)	167.75 ± 14.57	167.93 ± 20.22	177.80 ± 21.17	187.91 ± 13.71	0.0924
bFGF (count/field)	180.56 ± 14.51	184.99 ± 17.26	183.05 ± 21.49	194.52 ± 15.87	0.4367
PLGF (count/field)	167.44 ± 15.57	170.81 ± 21.73	173.23 ± 17.40	174.51 ± 16.75	0.7871

Abbreviations: PAS: Placenta accreta spectrum; ELISA: enzyme-linked immunosorbent assay; qRT-PCR: quantitative real-time polymerase chain reaction; IHC: immunohistochemistry; IGF-1: insulin-like growth factor-1; bFGF: basic fibroblast growth factor; PLGF: placental growth factor; BA: β-actin.

Note: All data presented in this table were expressed as means ± SD.

Data availability statement

All raw data that have been processed are owned by the first author and the corresponding author.